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Sample records for mexicana mexicana promastigotes

  1. Differential phagocytosis of Leishmania mexicana promastigotes and amastigotes by monocyte-derived dendritic cells.

    PubMed

    Argueta-Donohué, Jesús; Wilkins-Rodríguez, Arturo A; Aguirre-García, Magdalena; Gutiérrez-Kobeh, Laila

    2016-06-01

    Leishmania species are dimorphic protozoan parasites that live and replicate in the gut of sand flies as promastigotes or in mammalian hosts as amastigotes. Different immune cells, including DCs, and receptors differ in their involvement in phagocytosis of promastigotes and amastigotes and in recognition of different Leishmania species. In the case of L. mexicana, differences in phagocytosis of promastigotes and amastigotes by DCs and participation of C-type lectin receptors (CLRs) have not been established. In the present study, flow cytometry and confocal microscopy were used to investigate the phagocytosis by monocyte-derived dendritic cells (moDCs) of L. mexicana promastigotes and amastigotes in the presence or absence of immune serum during various periods of time. Blocking antibodies against mannose receptors and DC-SIGN were used to explore the participation of these receptors in the phagocytosis of L. mexicana by moDC. The major differences in interactions of L. mexicana promastigotes and amastigotes with moDC were found to occur within the first 3 hr, during which phagocytosis of promastigotes predominated as compared with opsonization of promastigotes and amastigotes. However, after 6 hr of incubation, opsonized promastigotes were preferentially phagocytosed as compared with unopsonized promastigotes and amastigotes and after 24 hr of incubation there were no differences in the phagocytosis of promastigotes and amastigotes. Finally, after 3 hr incubation, DC-SIGN was involved in the phagocytosis of promastigotes, but not of amastigotes. PMID:26399218

  2. Spectroscopic study of antileishmanial drug incubated in the promastigotes of Leishmania mexicana

    NASA Astrophysics Data System (ADS)

    Hung, J.; Castillo, J.; Jiménez, G.; Hasegawa, M.; Rodriguez, M.

    2003-11-01

    In this work we present spectroscopic study of Boldine (aporphine alkaloid) that possesses important biological activities, in particular, in interaction with the promastigotes of Leishmania mexicana. The results show the applicability of autofluorescence of this drug to determinate the possible mechanism of its biological action. The blue shift and hyperchromic effect in the emission spectrum of the drug in interaction with the parasite cells indicate an energy transference process between them. The morphological change of cell shape of the promastigotes treated with the drug is observed using confocal microscopy. This morphological cell-shape transformation evidences an important interaction between the drug studied and some protein of the parasite cell. Here we describe for the first time the fluorescence properties of the Boldine in the promastigotes of L. mexicana.

  3. Specific immunization of mice against Leishmania mexicana amazonensis using solubilized promastigotes

    NASA Technical Reports Server (NTRS)

    Barral-Netto, M.; Sadigursky, M.; Reed, S. G.; Sonnenfeld, G.

    1987-01-01

    In this work, it was demonstrated that mice (BALB/c strain) highly susceptible to Leishmania mexicana amazonensis can be protected against infection by this parasite by being preimmunized with whole solubilized (in a buffer that contained EDTA, NP-40, and SDS) promastigotes; the use of adjuvant or intact inactivated parasite cells is shown to be not necessary. The best immunization schedule consisted of three intravenous injections of 5 x 10 to the 7th parasite equivalents, administered one to eight weeks before infection. Immunized mice exhibited a marked inhibition of primary lesion development, reduced numbers of parasites in the spleen, and reduced death rate.

  4. Effect of ketoconazole on lethal action of amphotericin B on Leishmania mexicana promastigotes.

    PubMed Central

    Ramos, H; Saint-Pierre-Chazalet, M; Bolard, J; Cohen, B E

    1994-01-01

    The effect of ergosterol depletion by ketoconazole on the leishmanicidal activity of the pore-forming antibiotic amphotericin B (AmB) was investigated. Leishmania mexicana promastigotes were lysed within minutes by the addition of micromolar concentrations of AmB (0.5 microM) but became insensitive to AmB after growth in the presence of ketoconazole (0.25 microM, 90 h). Lipid chromatographic analysis indicated that under such conditions, ketoconazole depleted the major Leishmania sterols, dehydroepisterol and ergosterol. Plasma membrane vesicles prepared from ketoconazole-treated promastigotes exhibited a much reduced enhancement of their salt permeability after the addition of AmB at concentrations as high as 5 microM. This finding clearly indicates that upon ketoconazole treatment, the capacity of pore formation by the antibiotic is substantially impaired. The reduction of desmethyl sterols by ketoconazole was accompanied by a significant increase of 14-alpha-methyl sterols, but exogenous cholesterol remained unchanged. This ability of Leishmania promastigotes to incorporate cholesterol from the external medium may explain why ketoconazole-treated cells exhibited a much decreased but significative response to AmB when they were exposed to high AmB concentrations (2.5 or 5.0 microM). Parallel measurements by using a fluorescence energy transfer method indicated that binding of AmB to ketoconazole-treated Leishmania promastigotes and heat-transformed leishmanias was also decreased but to different extents, a finding that may be related to the differences in their sterol content. The results obtained clearly indicate that the specific interaction of AmB with desmethyl sterols, such as dehydroepisterol, ergosterol, and even exogenous cholesterol, is an absolute requirement for the lethal action exerted by this polyene antibiotic on L. mexicana promastigotes. PMID:8067741

  5. Expression of lipophosphoglycan, high-molecular weight phosphoglycan and glycoprotein 63 in promastigotes and amastigotes of Leishmania mexicana.

    PubMed

    Bahr, V; Stierhof, Y D; Ilg, T; Demar, M; Quinten, M; Overath, P

    1993-03-01

    The abundant surface glycoconjugate of Leishmania promastigotes, lipophosphoglycan (LPG), forms a blue-colored complex (lambda max = 649 nm) with the cationic dye Stains-all, which can be quantitated densitometrically on polyacrylamide gels of cell lysates. Promastigotes of Leishmania mexicana, Leishmania major and Leishmania donovani yield values of 1-3 x 10(6) LPG molecules cell-1. In amastigotes the LPG content is down-regulated below the detection limit (< 10(3) molecules cell-1) in L. mexicana and L. donovani, but remains significant in L. major (2 x 10(3) molecules cell-1). In the case of L. mexicana, these results are supported by immunological studies. Using several monoclonal and polyclonal antibodies, LPG is undetectable by immunoblotting in lysates of either amastigotes or infected macrophages and the amastigote surface is devoid of LPG as judged by immunofluorescence and immunoelectron microscopy. Immunoblotting experiments demonstrate that amastigotes synthesize hydrophilic high-molecular weight compounds which stain blue with Stains-all and cross-react with the monoclonal and polyvalent antibodies suggesting the presence of similar phosphoglycan structures as in LPG. The high-molecular weight phosphoglycan appears to be located in the lumen of the flagellar pocket of mouse lesion amastigotes and may be secreted from there into the lumen of the parasitophorous vacuole of parasitized macrophages. In L. mexicana promastigotes the surface protease gp63 is amphiphilic and comprises about 1% of the cellular proteins. In contrast, in amastigotes gp63-related proteins are predominantly hydrophilic; they amount to only about 0.1% of the cellular proteins and are mainly located in the lumen of the extended lysosomes (megasomes) characteristic for this species. PMID:8459823

  6. Characterization of polymer release from the flagellar pocket of Leishmania mexicana promastigotes.

    PubMed

    Stierhof, Y D; Ilg, T; Russell, D G; Hohenberg, H; Overath, P

    1994-04-01

    Trypanosomatids contain a unique compartment, the flagellar pocket, formed by an invagination of the plasma membrane at the base of the flagellum, which is considered to be the sole cellular site for endocytosis and exocytosis of macromolecules. The culture supernatant of Leishmania mexicana promastigotes, the insect stage of this protozoan parasite, contains two types of polymers: a filamentous acid phosphatase (sAP) composed of a 100-kD phosphoglycoprotein with non-covalently associated proteo high molecular weight phosphoglycan (proteo-HMWPG) and fibrous material termed network consisting of complex phosphoglycans. Secretion of both polymers is investigated using mAbs and a combination of light and electron microscopic techniques. Long filaments of sAP are detectable in the lumen of the flagellar pocket. Both sAP filaments and network material emerge from the ostium of the flagellar pocket. While sAP filaments detach from the cells, the fibrous network frequently remains associated with the anterior end of the parasites and can be found in the center of cell aggregates. The related species L. major forms similar networks. Since polymeric structures cannot be detected in intracellular compartments, it is proposed that monomeric or, possibly, oligomeric subunits synthesized in the cells are secreted into the flagellar pocket. Polymer formation from subunits is suggested to occur in the lumen of the pocket before release into the culture medium or, naturally, into the gut of infected sandflies. PMID:8163549

  7. Naturally azole-resistant Leishmania braziliensis promastigotes are rendered susceptible in the presence of terbinafine: comparative study with azole-susceptible Leishmania mexicana promastigotes.

    PubMed Central

    Rangel, H; Dagger, F; Hernandez, A; Liendo, A; Urbina, J A

    1996-01-01

    Leishmania braziliensis (isolate 2903) was naturally resistant to ketoconazole or the bis-triazole D0870, inhibitors of sterol C-14 demethylase, which produced only moderate effects on the proliferation of promastigotes at 10 microM. In contrast, Leishmania mexicana (isolate NR) was extremely susceptible to the azoles, as complete growth arrest and cell lysis were induced by incubation of the parasites with 0.05 microM concentrations of the drugs for 72 h. The opposite response was observed with terbinafine, an inhibitor of squalene epoxidase: L. braziliensis 2903 was three times more susceptible to the drug than L. mexicana NR (MICs of 5 and 15 microM, respectively). However, when the L. braziliensis stock was grown in the presence of 1 microM terbinafine, which by itself produced only marginal (< 10%) effects on growth, it became highly susceptible to the azoles, with an MIC of 0.03 microM. Analysis of cellular free sterols by high-resolution capillary gas chromatography coupled to mass spectrometry showed that 14-methyl sterols can support normal growth of L. braziliensis 2903 but not of L. mexicana NR. On the other hand, the higher susceptibility of the L. braziliensis isolate to terbinafine was correlated with a massive accumulation of squalene in the presence of the allylamine while no significant effects on L. mexicana sterol composition were observed at drug concentrations up to 1 microM. Thus, the > 300-fold increase in the susceptibility of L. braziliensis promastigotes to azoles in the presence of terbinafine was attributed to the combined effect of squalene and the methylated sterol precursors on the physical properties of the cell's membranes, leading to the loss of cell viability. Combination therapy with azoles and terbinafine in the treatment of human L. braziliensis infections deserves further study. PMID:9124841

  8. Leishmania mexicana: promastigotes and amastigotes secrete protein phosphatases and this correlates with the production of inflammatory cytokines in macrophages.

    PubMed

    Escalona-Montaño, A R; Ortiz-Lozano, D M; Rojas-Bernabé, A; Wilkins-Rodriguez, A A; Torres-Guerrero, H; Mondragón-Flores, R; Mondragón-Gonzalez, R; Becker, I; Gutiérrez-Kobeh, L; Aguirre-Garcia, M M

    2016-09-01

    Phosphatase activity of Leishmania spp. has been shown to deregulate the signalling pathways of the host cell. We here show that Leishmania mexicana promastigotes and amastigotes secrete proteins with phosphatase activity to the culture medium, which was higher in the Promastigote Secretion Medium (PSM) as compared with the Amastigote Secretion Medium (ASM) and was not due to cell lysis, since parasite viability was not affected by the secretion process. The biochemical characterization showed that the phosphatase activity present in PSM was higher in dephosphorylating the peptide END (pY) INASL as compared with the peptide RRA (pT)VA. In contrast, the phosphatase activity in ASM showed little dephosphorylating capacity for both peptides. Inhibition assays demonstrated that the phosphatase activity of both PSM and ASM was sensible only to protein tyrosine phosphatases inhibitors. An antibody against a protein phosphatase 2C (PP2C) of Leishmania major cross-reacted with a 44·9 kDa molecule in different cellular fractions of L. mexicana promastigotes and amastigotes, however, in PSM and ASM, the antibody recognized a protein about 70 kDa. By electron microscopy, the PP2C was localized in the flagellar pocket of amastigotes. PSM and ASM induced the production of tumor necrosis factor alpha, IL-1β, IL-12p70 and IL-10 in human macrophages. PMID:27220404

  9. Isolation and partial characterization of a broad specificity aminotransferase from Leishmania mexicana promastigotes.

    PubMed

    Vernal, J; Cazzulo, J J; Nowicki, C

    1998-10-30

    A broad specificity aminotransferase (BSAT), with high activity with both, aromatic amino acids and aspartate as substrates, was purified to homogeneity from promastigotes of Leishmania mexicana by a method involving chromatography on DEAE-cellulose, Red-120-Sepharose and Mono Q, and gel filtration on Sephacryl S-200. The purified enzyme showed a single band in SDS-polyacrylamide gel electrophoresis, with an apparent molecular mass of 45 kDa. Since the apparent molecular mass of the native enzyme, determined by gel filtration, was 90 kDa, the native enzyme is a dimer of similar subunits. The amino acid composition was determined, as well as the sequence of four internal peptides obtained by tryptic digestion. Two of these peptides, consisting of 49 amino acid residues in total, showed high similarity (57%) with corresponding sequences of plant aspartate aminotransferases, whereas they had only 33% identity with the aromatic aminotransferase of Escherichia coli, and 16% identity with the tyrosine aminotransferase from the related parasite Trypanosoma cruzi. The BSAT contained only one 1/2 Cys residue per monomer. The optimal pH for the enzyme reaction, with tyrosine and alpha-oxoglutarate as substrates, was 7.0. The apparent Km values for tyrosine, phenylalanine, tryptophan and glutamate, with oxaloacetate as co-substrate, were 1.3, 0.9, 0.9 and 171.8 mM, respectively; the value for aspartate with alpha-oxoglutarate as co-substrate was 2.5 mM, and that for alanine with alpha-oxoglutarate as co-substrate was 216 mM. The values for pyruvate, alpha-oxoglutarate and oxaloacetate, with tyrosine as co-substrate, were 5.6, 0.71 and 0.12 mM, respectively. These results suggest that the enzyme is a broad-specificity aminotransferase, able to transaminate the aromatic amino acids, aspartate, and to a lower extent alanine, with high sequence similarity to aspartate aminotransferases. PMID:9851609

  10. Leishmania mexicana promastigotes down regulate JNK and p-38 MAPK activation: Role in the inhibition of camptothecin-induced apoptosis of monocyte-derived dendritic cells.

    PubMed

    Rodríguez-González, Jorge; Wilkins-Rodríguez, Arturo; Argueta-Donohué, Jesús; Aguirre-García, Magdalena; Gutiérrez-Kobeh, Laila

    2016-04-01

    Dendritic cells (DC) are one of the principal host cells of the obligate intracellular parasite Leishmania. Inhibition of host cell apoptosis is a strategy employed by multiple pathogens to ensure their survival in the infected cell. We have previously shown that the infection of monocyte-derived dendritic cells (moDC) with Leishmania mexicana inhibits campthotecin-induced apoptosis. Nevertheless, the mechanisms involved in the inhibition of apoptosis of dendritic cells by Leishmania have not been established. Mitogen-activated protein kinases (MAPK) are key participants in the process of apoptosis and different species of Leishmania have been shown to regulate these kinases. In the present study, we analyzed the effect of L. mexicana promastigotes in the activation of JNK and p38 MAP kinase and their participation in the inhibition of apoptosis. The infection of moDC with L. mexicana promastigotes diminished significantly the phosphorylation of the MAP kinases JNK and p38. The inhibition of both kinases diminished DNA fragmentation, but in a major extent was the reduction of DNA fragmentation when JNK was inhibited. The capacity of L. mexicana promastigotes to diminish MAP kinases activation is probably one of the strategies employed to delay apoptosis induction in the infected moDC and may have implications for Leishmania pathogenesis by favoring the invasion of its host and the persistence of the parasite in the infected cells. PMID:26777406

  11. The glycoinositol phospholipids of Leishmania mexicana promastigotes. Evidence for the presence of three distinct pathways of glycolipid biosynthesis.

    PubMed

    McConville, M J; Collidge, T A; Ferguson, M A; Schneider, P

    1993-07-25

    Most macromolecules at the cell surface of parasitic protozoa of the genus Leishmania, including the major surface glycoproteins and a complex lipophosphoglycan (LPG), are attached to the plasma membrane via glycosyl-phosphatidylinositol (GPI) anchors. Free glycoinositol phospholipids (GIPLs) which are not linked to protein or phosphoglycan have also been found. In this study, we show that L. mexicana promastigotes synthesize two distinct GIPL lineages, comprising at least 10 glycolipid species. These structures were characterized using a combination of gas-liquid chromatography-mass spectrometry, methylation linkage analysis, and chemical and exoglycosidase sequencing. The major lineage contains GIPLs with the glycan structures Man alpha 1-3Man alpha 1-4GlcN (iM2), Man alpha 1-6(Man alpha 1-3)Man alpha 1-4GlcN (iM3), and Man alpha 1-2Man alpha 1-6(Man alpha 1-3)Man alpha 1-4GlcN (iM4), which are linked to alkylacyl-PI containing predominantly C16:0 and C18:0 fatty acids and C18:0 alkyl chains (referred to as the hybrid type GIPLs). A proportion of the iM3 and iM4 species (32 and 4%, respectively) are substituted with an ethanolamine-phosphate residue. The location of this residue on the core glucosamine residue was inferred from the results of methylation analyses and alpha-mannosidase digestion. The minor GIPL lineage contains GIPLs with the same glycan sequences as the glycolipid anchor of LPG (referred to as the type-2 GIPLs). The alkylacyl-PI or lyso-alkyl-PI lipid moieties of these GIPLs differ from those of the hybrid type GIPLs and from the main pool of alkylacyl-PI in containing significant levels of C24:0 and C26:0 alkyl chains. The most polar of these GIPLs, LPGp, has the properties expected of a biosynthetic precursor to the LPG, having the structure, [formula: see text] Finally, the GPI anchors of the major promastigote proteins were found to contain the glycan sequence Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcN, and an alkylacyl-PI lipid moiety which

  12. Proteophosphoglycans of Leishmania mexicana. Molecular cloning and characterization of the Leishmania mexicana ppg2 gene encoding the proteophosphoglycans aPPG and pPPG2 that are secreted by amastigotes and promastigotes.

    PubMed Central

    Göpfert, U; Goehring, N; Klein, C; Ilg, T

    1999-01-01

    Intracellular amastigotes of the pathogenic protozoon Leishmania mexicana secrete an extensively phosphoglycosylated proteophosphoglycan (aPPG) into the phagolysosome of mammalian host macrophages, that appears to fulfil important functions for the parasites. Promastigotes (the sandfly vector forms) of the same species secrete a proteophosphoglycan with identical protein backbone but exhibiting stage-specific phosphoglycosylation patterns [Klein, Göpfert, Goehring, Stierhof and Ilg (1999) Biochem. J. 344, 775-786]. In this study we report the cloning of the novel repeat-containing proteophosphoglycan gene ppg2 by antibody screening of a Leishmania mexicana amastigote cDNA expression library. ppg2 is equally expressed in promastigotes and amastigotes at the mRNA level. Targeted gene replacement of both alleles of the single copy gene ppg2 results in the loss of pPPG2 expression in promastigotes. Antisera against Escherichia coli-expressed ppg2 recognize the deglycosylated forms of aPPG as well as pPPG2. These results confirm that ppg2 encodes the protein backbones of aPPG and pPPG2. An unusual finding is that ppg2 exhibits two stable allelic forms, ppg2a and ppg2b. Their main difference lies in the number of central 72 bp DNA repeats (7 versus 8). ppg2a and ppg2b encode polypeptide chains of 574 and 598 amino acids, respectively, that show no homology to known proteins. The novel 24 amino acid Ser-rich peptide repeats encoded by the 72 bp DNA repeats are targets for Ser phosphoglycosylation in Leishmania mexicana. PMID:10585865

  13. Leishmania mexicana mexicana: quantitative analysis of the intracellular cycle.

    PubMed

    Doyle, P S; Engel, J C; Gam, A A; Dvorak, J A

    1989-12-01

    The complete intracellular cycle of the Leishmania mexicana mexicana G. S. strain was quantified in human macrophages and in the mouse IC-21 macrophage line utilizing a culture system that allows the direct observation of individual intracellular parasites. A wide range of pre-replicative lag periods exists, implying that promastigotes may be in any phase of their DNA synthetic cycle when phagocytosed by the macrophage. Amastigotes replicated 2-3 times, after which the host cell died and liberated amastigotes that were taken up by other macrophages and continued to replicate. The mean amastigote population-doubling time in human macrophages (17.5 h) was not statistically different from promastigotes growing in axenic culture (16.4 h), but was nearly 2-fold less than amastigotes growing in mouse-derived IC-21 macrophages (33.7 h). These observations are markedly different from cover-glass culture assays of Leishmania-macrophage interactions and provide an unambiguous description of the intracellular cycle of Leishmania mexicana mexicana. PMID:2608309

  14. Coordinate regulation of a family of promastigote-enriched mRNAs by the 3′UTR PRE element in Leishmania mexicana

    PubMed Central

    Holzer, Timothy R.; Mishra, Krishna K.; LeBowitz, Jonathan H.; Forney, James D.

    2009-01-01

    Post-transcriptional regulation is a key feature controlling gene expression in the protozoan parasite Leishmania. The nine-nucleotide paraflagellar rod regulatory element (PRE) in the 3′UTR of L. mexicana PFR2 is both necessary and sufficient for the observed ten-fold higher level of PFR2 mRNA in promastigotes compared to amastigotes. It is also found in the 3′UTRs of all known PFR genes. A search of the L. major Friedlin genomic database revealed several genes that share this cis element including a homolog of a heterotrimeric kinesin II subunit, and a gene that shares identity to a homolog of a Plasmodium antigen. In this study, we show that genes that harbor the PRE display promastigote-enriched transcript accumulation ranging from 4 – 15 fold. Northern analysis on episomal block substitution constructs revealed that the regulatory element is necessary for the proper steady-state accumulation of mRNA in L. mexicana paraflagellar rod gene 4 (PFR4). Also we show that the PRE plays a major role in the proper steady-state mRNA accumulation of PFR1, but may not account for the full regulatory mechanism acting on this mRNA. Our evidence suggests that the PRE coordinately regulates the mRNA abundance of not only the PFR family of genes, but in a larger group of genes that have unrelated functions. Although the PRE alone can regulate some mRNAs, it may also act in concert with additional elements to control other RNA transcripts. PMID:18023890

  15. Surface Zn-proteinase as a molecule for defense of Leishmania mexicana amazonensis promastigotes against cytolysis inside macrophage phagolysosomes.

    PubMed Central

    Seay, M B; Heard, P L; Chaudhuri, G

    1996-01-01

    The role of the surface membrane Zn-proteinase in protecting the cellular integrity of the macrophage parasite Leishmania mexicana amazonensis from intraphagolysosomal cytolysis was studied. These cells lose their infectivity to host macrophages after prolonged cultivation in axenic growth medium. The virulent and attenuated variants of the parasite cells were cloned. Failure of these attenuated parasite cells to survive inside macrophage phagolysosomes is associated with 20- to 50-fold reduction in the expression of surface gp63 protein. In situ inhibition of gp63 proteinase activity inside Leishmania-infected macrophage phagolysosomes with targeted delivery of an inhibitor of gp63 proteinase activity, 1,10-phenanthroline, selectively eliminated intracellular Leishmania amastigotes, further suggesting the importance of this proteinase in phagolysosomal survival of the parasite. An upstream sequence (US) of the gp63 gene was cloned in front of the bacterial chloramphenicol acetyltransferase (CAT) gene in plasmid pCATbasic. Transfection of L. mexicana amazonensis cells with this recombinant plasmid showed that expression of the CAT gene from this US is 15- to 20-fold higher in virulent clones than in avirulent clones of the parasite. Band shift analysis with the cloned US also showed that binding of protein(s) was 15- to 20-fold higher in virulent cell extract than in avirulent cell extract. Coating of attenuated cells or liposomes with proteolytically active gp63 protects them from degradation inside macrophage phagolysosomes. These results suggest a novel mechanism of survival of this phagolysosomal parasite with the help of its surface Zn-proteinase. PMID:8945556

  16. Photoacoustic monitoring of life cycles of Leishmania Mexicana

    NASA Astrophysics Data System (ADS)

    Arguello, C.; Acosta-Avalos, D.; Alvarado-Gil, J. J.; Vargas, H.

    1999-03-01

    Photoacoustic spectroscopy is used to monitor in situ, the difference between the two forms of the protozoan Leishmania Mexicana. Differences are the result of changes in the respiratory chain and could be attributed, according to our results, to the presence of cytochrome b in promastigotes and cytochrome c in amastigotes.

  17. Proteophosphoglycans of Leishmania mexicana. Identification, purification, structural and ultrastructural characterization of the secreted promastigote proteophosphoglycan pPPG2, a stage-specific glycoisoform of amastigote aPPG.

    PubMed Central

    Klein, C; Göpfert, U; Goehring, N; Stierhof, Y D; Ilg, T

    1999-01-01

    Protozoan parasites of the genus Leishmania secrete a range of proteophosphoglycans that appear to be important for successful colonization of the sandfly and for virulence in the mammalian host. A hallmark of these molecules is extensive phosphoglycosylation by phosphoglycan chains via the unusual linkage Manalpha1-PO(4)-Ser. In this study we have identified and purified to apparent homogeneity a novel proteophosphoglycan (pPPG2) which is secreted by Leishmania mexicana promastigotes (sandfly stage). Amino acid analysis and immunoblots using polypeptide-specific antisera suggest that pPPG2 shares a common protein backbone with a proteophosphoglycan (aPPG) secreted by Leishmania mexicana amastigotes (mammalian stage). Both pPPG2 and aPPG show a similar degree of Ser phosphoglycosylation (50. 5 mol% vs. 44.6 mol%), but the structure of their phosphoglycan chains is developmentally regulated: in contrast to aPPG which displays unique, complex and highly branched glycan chains [Ilg, Craik, Currie, Multhaup, and Bacic (1998) J. Biol. Chem. 273, 13509-13523], pPPG2 contains short unbranched structures consisting of >60 mol% neutral glycans, most likely (Manalpha1-2)(0-5)Man and Galbeta1-4Man, as well as about 40 mol% monophosphorylated glycans of the proposed structures PO(4)-6Galbeta1-4Man and PO(4)-6(Glcbeta1-3)Galbeta1-4Man. The major differences between pPPG2 and aPPG with respect to their apparent molecular mass, their ultrastructure and their proteinase sensitivity are most likely a consequence of this stage-specific glycosylation of their common protein backbone. PMID:10585864

  18. Proteophosphoglycans of Leishmania mexicana. Identification, purification, structural and ultrastructural characterization of the secreted promastigote proteophosphoglycan pPPG2, a stage-specific glycoisoform of amastigote aPPG.

    PubMed

    Klein, C; Göpfert, U; Goehring, N; Stierhof, Y D; Ilg, T

    1999-12-15

    Protozoan parasites of the genus Leishmania secrete a range of proteophosphoglycans that appear to be important for successful colonization of the sandfly and for virulence in the mammalian host. A hallmark of these molecules is extensive phosphoglycosylation by phosphoglycan chains via the unusual linkage Manalpha1-PO(4)-Ser. In this study we have identified and purified to apparent homogeneity a novel proteophosphoglycan (pPPG2) which is secreted by Leishmania mexicana promastigotes (sandfly stage). Amino acid analysis and immunoblots using polypeptide-specific antisera suggest that pPPG2 shares a common protein backbone with a proteophosphoglycan (aPPG) secreted by Leishmania mexicana amastigotes (mammalian stage). Both pPPG2 and aPPG show a similar degree of Ser phosphoglycosylation (50. 5 mol% vs. 44.6 mol%), but the structure of their phosphoglycan chains is developmentally regulated: in contrast to aPPG which displays unique, complex and highly branched glycan chains [Ilg, Craik, Currie, Multhaup, and Bacic (1998) J. Biol. Chem. 273, 13509-13523], pPPG2 contains short unbranched structures consisting of >60 mol% neutral glycans, most likely (Manalpha1-2)(0-5)Man and Galbeta1-4Man, as well as about 40 mol% monophosphorylated glycans of the proposed structures PO(4)-6Galbeta1-4Man and PO(4)-6(Glcbeta1-3)Galbeta1-4Man. The major differences between pPPG2 and aPPG with respect to their apparent molecular mass, their ultrastructure and their proteinase sensitivity are most likely a consequence of this stage-specific glycosylation of their common protein backbone. PMID:10585864

  19. Localized cutaneous leishmaniasis (chiclero's ulcer) in Mexico: sensitivity and specificity of ELISA for IgG antibodies to Leishmania mexicana mexicana.

    PubMed

    Garcia-Miss, M R; Andrade-Narvaez, F J; Esquivel-Viñas, R E; Simmonds-Diaz, E B; Canto-Lara, S B; Cruz-Ruiz, A L

    1990-01-01

    Sera from 223 human beings were assayed by enzyme-linked immunosorbent assay for immunoglobulin G antibodies to Leishmania mexicana mexicana promastigotes. 76% of 74 sera from patients with chiclero's ulcer gave positive reactions at a titre of 1:40; the positivity rate was lower in patients with long-standing infections. False positive reactions were obtained with only 7% of 56 sera from healthy individuals with positive Montenegro tests and with 6 of 7 sera from patients with Chagas disease. It is concluded that this method would be a useful contributory diagnostic method in cases of suspected chiclero's ulcer. PMID:2124389

  20. Immunoregulatory pathways in murine leishmaniasis: different regulatory control during Leishmania mexicana mexicana and Leishmania major infections.

    PubMed Central

    Alexander, J; Kaye, P M

    1985-01-01

    The effect of whole body sublethal gamma irradiation on the subsequent growth of Leishmania mexicana mexicana and Leishmania major was studied in CBA/Ca and BALB/c mice. Whereas BALB/c mice are highly susceptible to both parasites developing non healing progressively growing lesions at the site of cutaneous infection, CBA/Ca mice develop small healing cutaneous ulcers following subcutaneous infection with L. major but non healing lesions following subcutaneous infection with L.m. mexicana. Prior whole body sublethal irradiation of CBA/Ca mice, but not BALB/c mice, resulted in strong resistance against infection with L.m. mexicana: no lesions developed at the site of cutaneous infection. Irradiated BALB/c mice did, however, develop small lesions which healed when infected with L. major. The protective effects of irradiation coincided with the development of delayed type hypersensitivity. Both naive and sensitized nylon wool purified lymphocytes could restore susceptibility to L. major in irradiated BALB/c mice but only lymphocytes from long term infected donor mice adoptively transferred a non healing response to irradiated CBA/Ca mice infected with L.m. mexicana. Non-irradiated, L. major infected, CBA/Ca mice, but not similarly treated BALB/c mice, were found to be resistant to subsequent infection with L.m. mexicana. On the other hand, irradiated BALB/c mice infected with L. major were resistant to subsequent infectious challenge with L.m. mexicana. We suggest that the susceptibility of CBA/Ca mice to L.m. mexicana is under the control of an as yet unidentified gene which is not dependent on the generation of T suppressor cells and is bypassed by previous infection with L. major. Therefore, BALB/c mice immunized against L. major by prior sublethal irradiation are also resistant to L.m. mexicana. PMID:3907906

  1. Immunogenicity of a Salmonella typhi CVD 908 candidate vaccine strain expressing the major surface protein gp63 of Leishmania mexicana mexicana.

    PubMed

    González, C R; Noriega, F R; Huerta, S; Santiago, A; Vega, M; Paniagua, J; Ortiz-Navarrete, V; Isibasi, A; Levine, M M

    1998-01-01

    Attenuated Salmonella typhi are attractive for use as live vector vaccines to express protozoal antigens and deliver them to the human immune system. The gene encoding the mature form of Leishmania mexicana mexicana gp63 under control of tac promoter was integrated into the delta aroC locus of the chromosome of attenuated delta aroC, delta aroD S. typhi strain CVD 908. After oral immunization of BALB/c mice with two 1 x 10(9) colony forming unit doses given 21 days apart, CVD 908 omega (delta aroC::Ptac-gp63) elicited a broad T cell-mediated immune response against L. m. mexicana gp63 as demonstrated by: (1) lymphoproliferative response to fixed whole L. m. mexicana promastigotes; (2) activation of IL-2 (but not IL-4)-producing lymphocytes; (3) appearance of cytotoxic T cells against mouse mastocytoma cells expressing gp63. This T-cell mediated immune response was associated with significant protection in F1 (BALB/cXC57Bl/6) mice challenged in their footpads with a wild type strain of L. m. mexicana. PMID:9682357

  2. Lipophosphoglycan is not required for infection of macrophages or mice by Leishmania mexicana

    PubMed Central

    Ilg, Thomas

    2000-01-01

    Cell surface lipophosphoglycan (LPG) is commonly regarded as a multifunctional Leishmania virulence factor required for survival and development of these parasites in mammals. In this study, the LPG biosynthesis gene lpg1 was deleted in Leishmania mexicana by targeted gene replacement. The resulting mutants are deficient in LPG synthesis but still display on their surface and secrete phosphoglycan-modified molecules, most likely in the form of proteophosphoglycans, whose expression appears to be up-regulated. LPG-deficient L.mexicana promastigotes show no significant differences to LPG-expressing parasites with respect to attachment to, uptake into and multiplication inside macrophages. Moreover, in Balb/c and C57/BL6 mice, LPG-deficient L.mexicana clones are at least as virulent as the parental wild-type strain and lead to lethal disseminated disease. The results demonstrate that at least L.mexicana does not require LPG for experimental infections of macrophages or mice. Leishmania mexicana LPG is therefore not a virulence factor in the mammalian host. PMID:10790362

  3. Lipophosphoglycan is not required for infection of macrophages or mice by Leishmania mexicana.

    PubMed

    Ilg, T

    2000-05-01

    Cell surface lipophosphoglycan (LPG) is commonly regarded as a multifunctional Leishmania virulence factor required for survival and development of these parasites in mammals. In this study, the LPG biosynthesis gene lpg1 was deleted in Leishmania mexicana by targeted gene replacement. The resulting mutants are deficient in LPG synthesis but still display on their surface and secrete phosphoglycan-modified molecules, most likely in the form of proteophosphoglycans, whose expression appears to be up-regulated. LPG-deficient L.mexicana promastigotes show no significant differences to LPG-expressing parasites with respect to attachment to, uptake into and multiplication inside macrophages. Moreover, in Balb/c and C57/BL6 mice, LPG-deficient L.mexicana clones are at least as virulent as the parental wild-type strain and lead to lethal disseminated disease. The results demonstrate that at least L. mexicana does not require LPG for experimental infections of macrophages or mice. Leishmania mexicana LPG is therefore not a virulence factor in the mammalian host. PMID:10790362

  4. Natural Sesquiterpene Lactones Induce Oxidative Stress in Leishmania mexicana

    PubMed Central

    Barrera, Patricia; Sülsen, Valeria P.; Lozano, Esteban; Rivera, Mónica; Beer, María Florencia; Tonn, Carlos; Martino, Virginia S.; Sosa, Miguel A.

    2013-01-01

    Leishmaniasis is a worldwide parasitic disease, caused by monoflagellate parasites of the genus Leishmania. In the search for more effective agents against these parasites, the identification of molecular targets has been attempted to ensure the efficiency of drugs and to avoid collateral damages on the host's cells. In this work, we have investigated some of the mechanisms of action of a group of natural sesquiterpene lactones that are effective against Leishmania mexicana mexicana promastigotes. We first observed that the antiproliferative effect of mexicanin I (Mxc), dehydroleucodine (DhL), psilostachyin (Psi), and, at lesser extent, psilostachyin C (Psi C) is blocked by 1.5 mM reduced glutathione. The reducing agent was also able to reverse the early effect of the compounds, suggesting that lactones may react with intracellular sulfhydryl groups. Moreover, we have shown that all the sesquiterpene lactones, except Psi C, significantly decreased the endogenous concentration of glutathione within the parasite. Consistent with these findings, the active sesquiterpene lactones increased between 2.7 and 5.4 times the generation of ROS by parasites. These results indicate that the induction of oxidative stress is at least one of the mechanisms of action of DhL, Mxc, and Psi on parasites while Psi C would act by another mechanism. PMID:23861697

  5. Genetic characterization of glucose transporter function in Leishmania mexicana

    PubMed Central

    Burchmore, Richard J. S.; Rodriguez-Contreras, Dayana; McBride, Kathleen; Barrett, Michael P.; Modi, Govind; Sacks, David; Landfear, Scott M.

    2003-01-01

    Both insect and mammalian life cycle stages of Leishmania mexicana take up glucose and express all three isoforms encoded by the LmGT glucose transporter gene family. To evaluate glucose transporter function in intact parasites, a null mutant line has been created by targeted disruption of the LmGT locus that encompasses the LmGT1, LmGT2, and LmGT3 genes. This Δlmgt null mutant exhibited no detectable glucose transport activity. The growth rate of the Δlmgt knockout in the promastigote stage was reduced to a rate comparable with that of WT cells grown in the absence of glucose. Δlmgt cells also exhibited dramatically reduced infectivity to macrophages, demonstrating that expression of LmGT isoforms is essential for viability of amastigotes. Furthermore, WT L. mexicana were not able to grow as axenic culture form amastigotes if glucose was withdrawn from the medium, implying that glucose is an essential nutrient in this life cycle stage. Expression of either LmGT2 or LmGT3, but not of LmGT1, in Δlmgt null mutants significantly restored growth as promastigotes, but only LmGT3 expression substantially rescued amastigote growth in macrophages. Subcellular localization of the three isoforms was investigated in Δlmgt cells expressing individual LmGT isoforms. Using anti-LmGT antiserum and GFP-tagged LmGT fusion proteins, LmGT2 and LmGT3 were localized to the cell body, whereas LmGT1 was localized specifically to the flagellum. These results establish that each glucose transporter isoform has distinct biological functions in the parasite. PMID:12651954

  6. Cytokine mRNA expression in Peromyscus yucatanicus (Rodentia: Cricetidae) infected by Leishmania (Leishmania) mexicana.

    PubMed

    Loria-Cervera, Elsy Nalleli; Sosa-Bibiano, Erika Ivett; Van Wynsberghe, Nicole Raymonde; Saldarriaga, Omar Abdul; Melby, Peter C; Andrade-Narvaez, Fernando Jose

    2016-07-01

    Peromyscus yucatanicus, the main reservoir of Leishmania (Leishmania) mexicana in the Yucatan peninsula of Mexico, reproduces clinical and histological pictures of LCL in human as well as subclinical infection. Thus, we used this rodent as a novel experimental model. In this work, we analyzed cytokine mRNA expression in P. yucatanicus infected with L. (L.) mexicana. Animals were inoculated with either 2.5×10(6) or 1×10(2) promastigotes and cytokine expressions were analyzed by real-time RT-PCR in skin at 4 and 12weeks post-infection (wpi). Independently of the parasite inoculum none of the infected rodents had clinical signs of LCL at 4wpi and all expressed high IFN-γ mRNA. All P. yucatanicus inoculated with 2.5×10(6) promastigotes developed signs of LCL at 12wpi while the mice inoculated with 1×10(2) remained subclinical. At that time, both IFN-γ and IL-10 were expressed in P. yucatanicus with clinical and subclinical infections. Expressions of TNF-α and IL-4 were significantly higher in clinical animals (2.5×10(6)) compared with subclinical ones (1×10(2)). High TGF-β expression was observed in P. yucatanicus with clinical signs when compared with healthy animals. Results suggested that the clinical course of L. (L.) mexicana infection in P. yucatanicus was associated with a specific local pattern of cytokine production at 12wpi. PMID:27155064

  7. Nitric oxide production by Peromyscus yucatanicus (Rodentia) infected with Leishmania (Leishmania) mexicana

    PubMed Central

    Loría-Cervera, Elsy Nalleli; Sosa-Bibiano, Erika Ivett; Villanueva-Lizama, Liliana Estefanía; Van Wynsberghe, Nicole Raymonde; Canto-Lara, Silvia Beatriz; Batún-Cutz, José Luis; Andrade-Narváez, Fernando José

    2013-01-01

    Peromyscus yucatanicus (Rodentia: Cricetidae) is a primary reservoir of Leishmania (Leishmania) mexicana (Kinetoplastida: Trypanosomatidae). Nitric oxide (NO) generally plays a crucial role in the containment and elimination of Leishmania. The aim of this study was to determine the amount of NO produced by P. yucatanicus infected with L. (L.) mexicana. Subclinical and clinical infections were established in P. yucatanicus through inoculation with 1 x 102 and 2.5 x 106 promastigotes, respectively. Peritoneal macrophages were cultured alone or co-cultured with lymphocytes with or without soluble Leishmania antigen. The level of NO production was determined using the Griess reaction. The amount of NO produced was significantly higher (p ≤ 0.0001) in co-cultured macrophages and lymphocytes than in macrophages cultured alone. No differences in NO production were found between P. yucatanicus with subclinical L. (L.) mexicana infections and animals with clinical infections. These results support the hypothesis that the immunological mechanisms of NO production in P. yucatanicus are similar to those described in mouse models of leishmaniasis and, despite NO production, P. yucatanicus is unable to clear the parasite infection. PMID:23579796

  8. Leishmania (L.) mexicana Infected Bats in Mexico: Novel Potential Reservoirs

    PubMed Central

    Berzunza-Cruz, Miriam; Rodríguez-Moreno, Ángel; Gutiérrez-Granados, Gabriel; González-Salazar, Constantino; Stephens, Christopher R.; Hidalgo-Mihart, Mircea; Marina, Carlos F.; Rebollar-Téllez, Eduardo A.; Bailón-Martínez, Dulce; Balcells, Cristina Domingo; Ibarra-Cerdeña, Carlos N.; Sánchez-Cordero, Víctor; Becker, Ingeborg

    2015-01-01

    Leishmania (Leishmania) mexicana causes cutaneous leishmaniasis, an endemic zoonosis affecting a growing number of patients in the southeastern states of Mexico. Some foci are found in shade-grown cocoa and coffee plantations, or near perennial forests that provide rich breeding grounds for the sand fly vectors, but also harbor a variety of bat species that live off the abundant fruits provided by these shade-giving trees. The close proximity between sand flies and bats makes their interaction feasible, yet bats infected with Leishmania (L.) mexicana have not been reported. Here we analyzed 420 bats from six states of Mexico that had reported patients with leishmaniasis. Tissues of bats, including skin, heart, liver and/or spleen were screened by PCR for Leishmania (L.) mexicana DNA. We found that 41 bats (9.77%), belonging to 13 species, showed positive PCR results in various tissues. The infected tissues showed no evidence of macroscopic lesions. Of the infected bats, 12 species were frugivorous, insectivorous or nectarivorous, and only one species was sanguivorous (Desmodus rotundus), and most of them belonged to the family Phyllostomidae. The eco-region where most of the infected bats were caught is the Gulf Coastal Plain of Chiapas and Tabasco. Through experimental infections of two Tadarida brasiliensis bats in captivity, we show that this species can harbor viable, infective Leishmania (L.) mexicana parasites that are capable of infecting BALB/c mice. We conclude that various species of bats belonging to the family Phyllostomidae are possible reservoir hosts for Leishmania (L.) mexicana, if it can be shown that such bats are infective for the sand fly vector. Further studies are needed to determine how these bats become infected, how long the parasite remains viable inside these potential hosts and whether they are infective to sand flies to fully evaluate their impact on disease epidemiology. PMID:25629729

  9. Leishmania (L.) mexicana infected bats in Mexico: novel potential reservoirs.

    PubMed

    Berzunza-Cruz, Miriam; Rodríguez-Moreno, Ángel; Gutiérrez-Granados, Gabriel; González-Salazar, Constantino; Stephens, Christopher R; Hidalgo-Mihart, Mircea; Marina, Carlos F; Rebollar-Téllez, Eduardo A; Bailón-Martínez, Dulce; Balcells, Cristina Domingo; Ibarra-Cerdeña, Carlos N; Sánchez-Cordero, Víctor; Becker, Ingeborg

    2015-01-01

    Leishmania (Leishmania) mexicana causes cutaneous leishmaniasis, an endemic zoonosis affecting a growing number of patients in the southeastern states of Mexico. Some foci are found in shade-grown cocoa and coffee plantations, or near perennial forests that provide rich breeding grounds for the sand fly vectors, but also harbor a variety of bat species that live off the abundant fruits provided by these shade-giving trees. The close proximity between sand flies and bats makes their interaction feasible, yet bats infected with Leishmania (L.) mexicana have not been reported. Here we analyzed 420 bats from six states of Mexico that had reported patients with leishmaniasis. Tissues of bats, including skin, heart, liver and/or spleen were screened by PCR for Leishmania (L.) mexicana DNA. We found that 41 bats (9.77%), belonging to 13 species, showed positive PCR results in various tissues. The infected tissues showed no evidence of macroscopic lesions. Of the infected bats, 12 species were frugivorous, insectivorous or nectarivorous, and only one species was sanguivorous (Desmodus rotundus), and most of them belonged to the family Phyllostomidae. The eco-region where most of the infected bats were caught is the Gulf Coastal Plain of Chiapas and Tabasco. Through experimental infections of two Tadarida brasiliensis bats in captivity, we show that this species can harbor viable, infective Leishmania (L.) mexicana parasites that are capable of infecting BALB/c mice. We conclude that various species of bats belonging to the family Phyllostomidae are possible reservoir hosts for Leishmania (L.) mexicana, if it can be shown that such bats are infective for the sand fly vector. Further studies are needed to determine how these bats become infected, how long the parasite remains viable inside these potential hosts and whether they are infective to sand flies to fully evaluate their impact on disease epidemiology. PMID:25629729

  10. Gluconeogenesis in Leishmania mexicana: contribution of glycerol kinase, phosphoenolpyruvate carboxykinase, and pyruvate phosphate dikinase.

    PubMed

    Rodriguez-Contreras, Dayana; Hamilton, Nicklas

    2014-11-21

    Gluconeogenesis is an active pathway in Leishmania amastigotes and is essential for their survival within the mammalian cells. However, our knowledge about this pathway in trypanosomatids is very limited. We investigated the role of glycerol kinase (GK), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate phosphate dikinase (PPDK) in gluconeogenesis by generating the respective Leishmania mexicana Δgk, Δpepck, and Δppdk null mutants. Our results demonstrated that indeed GK, PEPCK, and PPDK are key players in the gluconeogenesis pathway in Leishmania, although stage-specific differences in their contribution to this pathway were found. GK participates in the entry of glycerol in promastigotes and amastigotes; PEPCK participates in the entry of aspartate in promastigotes, and PPDK is involved in the entry of alanine in amastigotes. Furthermore, the majority of alanine enters into the pathway via decarboxylation of pyruvate in promastigotes, whereas pathway redundancy is suggested for the entry of aspartate in amastigotes. Interestingly, we also found that l-lactate, an abundant glucogenic precursor in mammals, was used by Leishmania amastigotes to synthesize mannogen, entering the pathway through PPDK. On the basis of these new results, we propose a revision in the current model of gluconeogenesis in Leishmania, emphasizing the differences between amastigotes and promastigotes. This work underlines the importance of studying the trypanosomatid intracellular life cycle stages to gain a better understanding of the pathologies caused in humans. PMID:25288791

  11. β-Adrenergic blockade protects BALB/c mice against infection with a small inoculum of Leishmania mexicana mexicana (LV4).

    PubMed

    García-Miss, María del R; Mut-Martín, Mirza C; Góngora-Alfaro, José L

    2015-01-01

    In order to test the influence of the sympathetic nervous system on Leishmania mexicana infection, groups of female BALB/c mice were treated (i.p.) with the non-selective β-adrenergic receptor (β-AR) antagonist (S)-propranolol (5mg/kg thrice a day), the β2-AR agonist clenbuterol (1mg/kg once a day) or the α2-AR antagonist yohimbine (2mg/kg twice a day) during 5days. During the second day of treatments, mice were inoculated in the footpad with 1×10(6) or 1×10(3) metacyclic promastigotes of L. mexicana mexicana (LV4). The lesion size was measured weekly, and parasite burden on week 12. In mice treated with (S)-propranolol, the percentage of splenic T lymphocytes producing IFN-γ after antigen challenge was determined by flow cytometry. In mice infected with 1×10(6) parasites, only (S)-propranolol caused a reduction of footpad swelling (p<0.05, weeks 11-12), without effects on parasite burden, or in the percentage of IFN-γ-immunopositive CD4(+) or CD8(+) T lymphocytes. In mice infected with 1×10(3) parasites, the effects of treatments vs. control group were as follows: (a) inhibition of footpad swelling by (S)-propranolol (p<0.01, weeks 3-12), clenbuterol (p<0.05, weeks 7-10), and yohimbine (p<0.01, week 7); (b) a decrease of the parasite burden by (S)-propranolol (p<0.01) and yohimbine (p<0.05); (c) in control mice the percentage of CD4(+) T-cells producing IFN-γ was 6.2±0.5%, while in those treated with (S)-propranolol it increased to 8.7±0.6% (p<0.01); (d) in control mice the percentage of CD8(+) T-cells producing IFN-γ was 3.1±0.4%, while in those treated with (S)-propranolol it increased to 10.4±0.2% (p<0.01). These results indicate that the blockade of β-ARs during infection of BALB/c mice with an inoculum of L. mexicana mexicana similar to that delivered by the bite of a sand fly produces a Th1 bias in the immune response, favoring an increment of T lymphocytes secreting IFN-γ, which correlated with a reduced parasite burden (p<0.05, Spearman

  12. Dronedarone, an Amiodarone Analog with Improved Anti-Leishmania mexicana Efficacy

    PubMed Central

    Casanova, Paola; Hernandez-Rodriguez, Vanessa; Mujica-Gonzalez, Sheira; Parra-Gimenez, Nereida; Plaza-Rojas, Lourdes; Concepcion, Juan Luis; Liu, Yi-Liang; Oldfield, Eric; Paniz-Mondolfi, Alberto; Suarez, Alirica I.

    2014-01-01

    Dronedarone and amiodarone are cationic lipophilic benzofurans used to treat cardiac arrhythmias. They also have activity against the parasitic protozoan Trypanosoma cruzi, the causative agent of Chagas' disease. They function by disrupting intracellular Ca2+ homeostasis of the parasite and by inhibiting membrane sterol (ergosterol) biosynthesis. Amiodarone also has activity against Leishmania mexicana, suggesting that dronedarone might likewise be active against this organism. This might be of therapeutic interest, since dronedarone is thought to have fewer side effects in humans than does amiodarone. We show here that dronedarone effectively inhibits the growth of L. mexicana promastigotes in culture and, more importantly, has excellent activity against amastigotes inside infected macrophages (the clinically relevant form) without affecting the host cell, with the 50% inhibitory concentrations against amastigotes being 3 orders of magnitude lower than those obtained previously with T. cruzi amastigotes (0.65 nM versus 0.75 μM). As with amiodarone, dronedarone affects intracellular Ca2+ homeostasis in the parasite, inducing an elevation of intracellular Ca2+ levels. This is achieved by rapidly collapsing the mitochondrial membrane potential and inducing an alkalinization of acidocalcisomes at a rate that is faster than that observed with amiodarone. We also show that dronedarone inhibits parasite oxidosqualene cyclase, a key enzyme in ergosterol biosynthesis known to be vital for survival. Overall, our results suggest the possibility of repurposing dronedarone as a treatment for cutaneous, and perhaps other, leishmaniases. PMID:24492373

  13. Dronedarone, an amiodarone analog with improved anti-Leishmania mexicana efficacy.

    PubMed

    Benaim, Gustavo; Casanova, Paola; Hernandez-Rodriguez, Vanessa; Mujica-Gonzalez, Sheira; Parra-Gimenez, Nereida; Plaza-Rojas, Lourdes; Concepcion, Juan Luis; Liu, Yi-Liang; Oldfield, Eric; Paniz-Mondolfi, Alberto; Suarez, Alirica I

    2014-01-01

    Dronedarone and amiodarone are cationic lipophilic benzofurans used to treat cardiac arrhythmias. They also have activity against the parasitic protozoan Trypanosoma cruzi, the causative agent of Chagas' disease. They function by disrupting intracellular Ca2+ homeostasis of the parasite and by inhibiting membrane sterol (ergosterol) biosynthesis. Amiodarone also has activity against Leishmania mexicana, suggesting that dronedarone might likewise be active against this organism. This might be of therapeutic interest, since dronedarone is thought to have fewer side effects in humans than does amiodarone. We show here that dronedarone effectively inhibits the growth of L. mexicana promastigotes in culture and, more importantly, has excellent activity against amastigotes inside infected macrophages (the clinically relevant form) without affecting the host cell, with the 50% inhibitory concentrations against amastigotes being 3 orders of magnitude lower than those obtained previously with T. cruzi amastigotes (0.65 nM versus 0.75 μM). As with amiodarone, dronedarone affects intracellular Ca2+ homeostasis in the parasite, inducing an elevation of intracellular Ca2+ levels. This is achieved by rapidly collapsing the mitochondrial membrane potential and inducing an alkalinization of acidocalcisomes at a rate that is faster than that observed with amiodarone. We also show that dronedarone inhibits parasite oxidosqualene cyclase, a key enzyme in ergosterol biosynthesis known to be vital for survival. Overall, our results suggest the possibility of repurposing dronedarone as a treatment for cutaneous, and perhaps other, leishmaniases. PMID:24492373

  14. In vitro activity of synthetic tetrahydroindeno[2,1-c]quinolines on Leishmania mexicana.

    PubMed

    Hernández-Chinea, Concepción; Carbajo, Erika; Sojo, Felipe; Arvelo, Francisco; Kouznetsov, Vladimir V; Romero-Bohórquez, Arnold R; Romero, Pedro J

    2015-12-01

    New synthetic compounds based on tetrahydroindenoquinoline structure were evaluated for their in vitro antileishmanial activities. The seven compounds assayed have antiproliferative activities against promastigotes of Leishmania mexicana. Compound 1 and 3 were the most active (IC50 1.0 μg/ml) and showed high selectivity towards the parasite. These compounds were selected to evaluate their effect on promastigote morphology and mitochondrial transmembrane potential as well as on the amastigote capability to survive into macrophages J774 cell line. Whereas compound 1 affected the promastigote cell cycle, compound 3 induced morphological changes and the total collapse of the mitochondrial transmembrane potential, a hallmark of apoptosis. Both compounds also affected the amastigote form of the parasite, decreasing their survival rate in J774 macrophages. Due to the greatest selectivity index, the apparent effect as apoptotic inducer and its sustained inhibition on intracellular amastigote replication, compound 3 is the best candidate to be tested in vivo. This compound is worth considering for the development of new antileishmanial drugs. PMID:26148815

  15. Purification, partial characterization and immunolocalization of a proteophosphoglycan secreted by Leishmania mexicana amastigotes.

    PubMed

    Ilg, T; Stierhof, Y D; McConville, M J; Overath, P

    1995-02-01

    The intracellular amastigote form of the parasitic protozoon Leishmania mexicana expresses a high-molecular weight phosphoglycan, which is antigenically related to the surface glycolipid lipophosphoglycan and the secreted enzyme acid phosphatase of Leishmania promastigotes. This antigen was purified from a cell-free homogenate of infected mouse tissue and from amastigotes. Compositional and immunological analysis of the purified components indicate a proteophosphoglycan structure consisting of serine-rich polypeptide chains and mild acid-labile phosphooligosaccharides capped by mannooligosaccharides. Immunofluorescence and immunoelectron microscopy of parasitized mouse peritoneal macrophages and infected mouse tissue suggest that the proteophosphoglycan is secreted in large amounts by amastigotes via their flagellar pockets into the parasitophorous vacuoles of host cells. In some infected macrophages proteophosphoglycan is also located in vesicles apparently originating from the parasitophorous vacuole, which demonstrates redistribution of a secreted amastigote antigen in parasitized host cells. PMID:7774606

  16. Homology modeling of LmxMPK4 of Leishmania mexicana and virtual screening of potent inhibitors against it.

    PubMed

    Gupta, Chhedi Lal; Khan, Mohd Kalim Ahmad; Khan, Mohd Faheem; Tiwari, Ashok K

    2013-06-01

    Leishmaniasis is one of the most important diseases of mankind. In the life cycle of Leishmania mexicana, two most important developmental stages are observed. In insect vector it is in promastigote form and in mammalian macrophages is the amastigote form. The family of protein kinases are extremely important regulators of many different cellular processes such as transcriptional control, cell cycle development and differentiation, and also draw much attention as possible drug targets for protozaon parasites. Leishmania mexicana mitogen activated protein kinase 4 (LmxMPK4) is essential for proliferation and survival of the parasite promastigote and amastigote forms and is a potential drug target for leishmaniasis. The existing therapy for leishmaniasis is not enough due to host toxicity and drug resistance. The experimental 3D structure of this protein has not yet been determined. In this study, we have used homology modelling techniques to generate the 3D structure of LmxMPK4 and selected effective inhibitors by ZINC database on the basis of structure of berberine alkaloid for molecular docking studies with LmxMPK4. The inhibitors ZINC05999210, ZINC40402312 and ZINC40977377 were found to be more potent for inhibition of leishmaniasis due to the robust binding affinity and strong inhibition constant (Ki) of the protein-ligand interactions. This finding may help to understand the nature of MAP kinase and development of specific anti-leishmanial therapies. PMID:23740395

  17. Regulatory volume decrease in Leishmania mexicana: effect of anti-microtubule drugs.

    PubMed

    Dagger, Francehuli; Valdivieso, Elizabeth; Marcano, Ana K; Ayesta, Carlos

    2013-02-01

    The trypanosomatid cytoskeleton is responsible for the parasite's shape and it is modulated throughout the different stages of the parasite's life cycle. When parasites are exposed to media with reduced osmolarity, they initially swell, but subsequently undergo compensatory shrinking referred to as regulatory volume decrease (RVD). We studied the effects of anti-microtubule (Mt) drugs on the proliferation of Leishmania mexicana promastigotes and their capacity to undergo RVD. All of the drugs tested exerted antiproliferative effects of varying magnitudes [ansamitocin P3 (AP3)> trifluoperazine > taxol > rhizoxin > chlorpromazine]. No direct relationship was found between antiproliferative drug treatment and RVD. Similarly, Mt stability was not affected by drug treatment. Ansamitocin P3, which is effective at nanomolar concentrations, blocked amastigote-promastigote differentiation and was the only drug that impeded RVD, as measured by light dispersion. AP3 induced 2 kinetoplasts (Kt) 1 nucleus cells that had numerous flagella-associated Kts throughout the cell. These results suggest that the dramatic morphological changes induced by AP3 alter the spatial organisation and directionality of the Mts that are necessary for the parasite's hypotonic stress-induced shape change, as well as its recovery. PMID:23440120

  18. Inhibition profile of Leishmania mexicana arginase reveals differences with human arginase I

    PubMed Central

    Riley, Eric; Roberts, Sigrid C.; Ullman, Buddy

    2011-01-01

    Arginase (ARG), the enzyme that catalyzes the conversion of arginine to ornithine and urea, is the first and committed step in polyamine biosynthesis in Leishmania. The creation of a conditionally lethal Δarg null mutant in Leishmania mexicana has established that ARG is an essential enzyme for the promastigote form of the parasite and that the enzyme provides an important defense mechanism for parasite survival in the eukaryotic host. Furthermore, human ARGI (HsARGI) has also been implicated as a key factor in parasite proliferation. Thus, inhibitors of ARG offer a rational paradigm for drug design. To initiate a search for inhibitors of the L. mexicana ARG (LmARG), recombinant LmARG and HsARGI enzymes were purified from Escherichia coli. Both LmARG and HsARGI were specific for L-arginine and exhibited no activity with either D-arginine or agmatine as possible substrates. LmARG exhibited a Km of 25 ± 4 mM for L-arginine, a pH optimum ~9.0, and was dependent upon the presence of a divalent cation, preferentially manganese. A Km of 13.5 ± 2 mM for L-arginine was calculated for the HsARGI. A collection of 37 compounds was evaluated against both enzymes. Twelve of these compounds were identified as being either strong inhibitors of both LmARG and HsARGI or differential inhibitors between the two enzymes. Of the 12 compounds, six were selected for further analysis and the type and extent of inhibition determined. PMID:21232540

  19. BIOADHESION IN CAULERPA MEXICANA (CHLOROPHYTA): RHIZOID-SUBSTRATE ADHESION(1).

    PubMed

    Fagerberg, Wayne R; Towle, Jennifer; Dawes, Clinton J; Böttger, Anne

    2012-04-01

    The attachment of the psammophytic alga Caulerpa mexicana Sond. ex Kütz., a coenocytic green alga, to crushed CaCO3 particles was examined utilizing the scanning electron microscope and fluorescently tagged antivitronectin antibodies. Plants attached to the substrate through morphologically variable tubular rhizoidal extensions that grew from the stolon. In this study, we describe two means of attachment: (i) the rhizoid attachment to limestone gravel by thigmoconstriction, where tubular extensions of the rhizoid wrapped tightly around the substrate and changed morphology to fit tightly into crevices in the limestone, and (ii) through adhesion pads that formed in contact with the limestone granules. Flattened rhizoidal pads were observed to secrete a fibrillar material that contained vitronectin-like proteins identified through immunolocialization and that facilitated binding of the rhizoid to the substrate. PMID:27009715

  20. Pentalinon andrieuxii root extract is effective in the topical treatment of cutaneous leishmaniasis caused by Leishmania mexicana

    PubMed Central

    Lezama-Dávila, Claudio M.; Pan, Li; Isaac-Márquez, Angelica P.; Terrazas, Cesar; Oghumu, Steve; Isaac-Márquez, Ricardo; Pech-Dzib, MY; Barbi, Joseph; Calomeni, Edward; Parinandi, Narasimham; Kinghorn, A. Douglas; Satoskar, Abhay R.

    2014-01-01

    Cutaneous leishmaniasis (CL) manifests as localized skin lesions, which lead to significant tissue destruction and disfigurement. In the Yucatan Peninsula, Mayan traditional healers use Pentalinon andrieuxii Muell.-Arg. (Apocynaceae) roots for the topical treatment of CL. Here, we studied the effect of P. andrieuxii root hexane extract (PARE) on the parasites and host cells in vitro and examined its efficacy in the topical treatment of CL caused by L. mexicana. PARE exhibited potent antiparasitic activity in vitro against promastigotes as well as amastigotes residing in macrophages. Electron microscopy of PARE-treated parasites revealed direct membrane damage. PARE also activated NF-κB and enhanced IFN-γR and MHC class II expression and TNF-α production in macrophages. In addition, PARE induced production of the Th1 promoting cytokine IL-12 in dendritic cells as well as enhanced expression of the co-stimulatory molecules CD40, CD80 and CD86. In vivo studies showed that L. mexicana-infected mice treated by topical application of PARE resulted in the significant reduction in lesion size and parasite burden compared to controls. These findings indicate that PARE could be used as an alternative therapy for the topical treatment of CL. PMID:24347110

  1. Pentalinon andrieuxii root extract is effective in the topical treatment of cutaneous leishmaniasis caused by Leishmania mexicana.

    PubMed

    Lezama-Dávila, Claudio M; Pan, Li; Isaac-Márquez, Angelica P; Terrazas, Cesar; Oghumu, Steve; Isaac-Márquez, Ricardo; Pech-Dzib, M Y; Barbi, Joseph; Calomeni, Edward; Parinandi, Narasimham; Kinghorn, A Douglas; Satoskar, Abhay R

    2014-06-01

    Cutaneous leishmaniasis (CL) manifests as localized skin lesions, which lead to significant tissue destruction and disfigurement. In the Yucatan Peninsula, Mayan traditional healers use Pentalinon andrieuxii Muell.-Arg. (Apocynaceae) roots for the topical treatment of CL. Here, we studied the effect of P. andrieuxii root hexane extract (PARE) on the parasites and host cells in vitro and examined its efficacy in the topical treatment of CL caused by Leishmania mexicana. PARE exhibited potent antiparasitic activity in vitro against promastigotes as well as amastigotes residing in macrophages. Electron microscopy of PARE-treated parasites revealed direct membrane damage. PARE also activated nuclear factor kappaB and enhanced interferon-γ receptor and MHC class II expression and TNF-α production in macrophages. In addition, PARE induced production of the Th1 promoting cytokine IL-12 in dendritic cells as well as enhanced expression of the co-stimulatory molecules CD40, CD80, and CD86. In vivo studies showed that L. mexicana-infected mice treated by topical application of PARE resulted in the significant reduction in lesion size and parasite burden compared to controls. These findings indicate that PARE could be used as an alternative therapy for the topical treatment of CL. PMID:24347110

  2. Regulation and biological function of a flagellar glucose transporter in Leishmania mexicana: a potential glucose sensor.

    PubMed

    Rodriguez-Contreras, Dayana; Aslan, Hamide; Feng, Xiuhong; Tran, Khoa; Yates, Phillip A; Kamhawi, Shaden; Landfear, Scott M

    2015-01-01

    In Leishmania mexicana parasites, a unique glucose transporter, LmxGT1, is selectively targeted to the flagellar membrane, suggesting a possible sensory role that is often associated with ciliary membrane proteins. Expression of LmxGT1 is down-regulated ∼20-fold by increasing cell density but is up-regulated ∼50-fold by depleting glucose from the medium, and the permease is strongly down-regulated when flagellated insect-stage promastigotes invade mammalian macrophages and transform into intracellular amastigotes. Regulation of LmxGT1 expression by glucose and during the lifecycle operates at the level of protein stability. Significantly, a ∆lmxgt1 null mutant, grown in abundant glucose, undergoes catastrophic loss of viability when parasites deplete glucose from the medium, a property not exhibited by wild-type or add-back lines. These results suggest that LmxGT1 may function as a glucose sensor that allows parasites to enter the stationary phase when they deplete glucose and that in the absence of this sensor, parasites do not maintain viability when they run out of glucose. However, alternate roles for LmxGT1 in monitoring glucose availability are considered. The absence of known sensory receptors with defined ligands and biologic functions in Leishmania and related kinetoplastid parasites underscores the potential significance of these observations. PMID:25300620

  3. The Mexican Seismic Network (Red Sísmica Mexicana)

    NASA Astrophysics Data System (ADS)

    Valdes-Gonzales, C. M.; Arreola-Manzano, J.; Castelan-Pescina, G.; Alonso-Rivera, P.; Saldivar-Rangel, M. A.; Rodriguez-Arteaga, O. O.; Lopez-Lena-Villasana, R.

    2014-12-01

    The Mexican Seismic Network (Red Sísmica Mexicana) was created to give sufficient information and opportune to make decisions in order to mitigate seismic and tsunami risk. This was a Mexican government initiative headed by CENAPRED (National Disaster Prevention Center) who made an effort to integrated academic institutions and civil agencies to work together through a collaboration agreement. This network is supported by Universidad National Autónoma de México (UNAM) and its seismic networks (Broad Band and Strong Motion), the Centro de Instrumentación y Registro Sismico (CIRES) with its Earthquake Early Warning System that covers the Guerrero Gap and Oaxaca earthquakes, The Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE) with the support of its expertise in tsunami observation and the Secretaria de Marina (SEMAR) to monitor the sea level and operate the Mexican Tsunami Warning Center. The institutions involved in this scope have the compromise to interchange and share the data and advice to the Civil Protection authorities.

  4. Glycosomal membrane proteins and lipids from Leishmania mexicana.

    PubMed

    Quiñones, Wilfredo; Cáceres, Ana J; Ruiz, Maria Tibisay; Concepción, Juan Luis

    2015-04-01

    Constituents of the glycosomal membrane from Leishmania mexicana should play a critical role in the coordination of metabolic processes occurring in the cytosol and those compartmentalized within glycosomes. We have made an inventory of glycosomal membrane-associated proteins using approaches specific for enriching both integral and peripheral membrane proteins. Surprisingly, 70% of the proteins were recovered in the hydrophobic fraction of membranes solubilized with Triton X-114, while 20% were present in the soluble fraction obtained upon treatment with Na2CO3. 14 major polypeptides, ranging in molecular weight from 65 to 16 kDa, were found to be associated with the membrane, nine of them behaving as integral membrane proteins. Assessment of their topology in the membrane indicated that the polypeptides of 56, 50, 46 and 32 kDa have no domains exposed to the cytosol. The 50 kDa protein is the most abundant one of the glycosomal membrane, where it is peripherically located at the matrix face. The major phospholipids of glycosomal membranes are phosphatidyl-ethanolamine, phosphatidyl-choline and phosphatidyl-serine, with smaller proportions of sphingomyelin and phosphatidyl-inositol. The sterols found were of 5-dehydroepisterol, ergosta-5,7,24(24(1))-trien-3β-ol, and also their precursors, consistent with the notion that these organelles are involved in de novo biosynthesis of sterols in trypanosomatids. PMID:25499533

  5. Comparative Life Cycle Transcriptomics Revises Leishmania mexicana Genome Annotation and Links a Chromosome Duplication with Parasitism of Vertebrates

    PubMed Central

    Fiebig, Michael; Kelly, Steven; Gluenz, Eva

    2015-01-01

    Leishmania spp. are protozoan parasites that have two principal life cycle stages: the motile promastigote forms that live in the alimentary tract of the sandfly and the amastigote forms, which are adapted to survive and replicate in the harsh conditions of the phagolysosome of mammalian macrophages. Here, we used Illumina sequencing of poly-A selected RNA to characterise and compare the transcriptomes of L. mexicana promastigotes, axenic amastigotes and intracellular amastigotes. These data allowed the production of the first transcriptome evidence-based annotation of gene models for this species, including genome-wide mapping of trans-splice sites and poly-A addition sites. The revised genome annotation encompassed 9,169 protein-coding genes including 936 novel genes as well as modifications to previously existing gene models. Comparative analysis of gene expression across promastigote and amastigote forms revealed that 3,832 genes are differentially expressed between promastigotes and intracellular amastigotes. A large proportion of genes that were downregulated during differentiation to amastigotes were associated with the function of the motile flagellum. In contrast, those genes that were upregulated included cell surface proteins, transporters, peptidases and many uncharacterized genes, including 293 of the 936 novel genes. Genome-wide distribution analysis of the differentially expressed genes revealed that the tetraploid chromosome 30 is highly enriched for genes that were upregulated in amastigotes, providing the first evidence of a link between this whole chromosome duplication event and adaptation to the vertebrate host in this group. Peptide evidence for 42 proteins encoded by novel transcripts supports the idea of an as yet uncharacterised set of small proteins in Leishmania spp. with possible implications for host-pathogen interactions. PMID:26452044

  6. Conditional gene deletion with DiCre demonstrates an essential role for CRK3 in L eishmania mexicana cell cycle regulation

    PubMed Central

    Duncan, Samuel M.; Myburgh, Elmarie; Philipon, Cintia; Brown, Elaine; Meissner, Markus; Brewer, James

    2016-01-01

    Summary Leishmania mexicana has a large family of cyclin‐dependent kinases (CDKs) that reflect the complex interplay between cell cycle and life cycle progression. Evidence from previous studies indicated that Cdc2‐related kinase 3 (CRK3) in complex with the cyclin CYC6 is a functional homologue of the major cell cycle regulator CDK1, yet definitive genetic evidence for an essential role in parasite proliferation is lacking. To address this, we have implemented an inducible gene deletion system based on a dimerised Cre recombinase (diCre) to target CRK3 and elucidate its role in the cell cycle of L. mexicana. Induction of diCre activity in promastigotes with rapamycin resulted in efficient deletion of floxed CRK3, resulting in G2/M growth arrest. Co‐expression of a CRK3 transgene during rapamycin‐induced deletion of CRK3 resulted in complementation of growth, whereas expression of an active site CRK3 T178E mutant did not, showing that protein kinase activity is crucial for CRK3 function. Inducible deletion of CRK3 in stationary phase promastigotes resulted in attenuated growth in mice, thereby confirming CRK3 as a useful therapeutic target and diCre as a valuable new tool for analyzing essential genes in Leishmania. PMID:26991545

  7. Kharon1 Null Mutants of Leishmania mexicana Are Avirulent in Mice and Exhibit a Cytokinesis Defect within Macrophages

    PubMed Central

    Sanchez, Marco A.; Valli, Jessica; Gluenz, Eva; Landfear, Scott M.

    2015-01-01

    In a variety of eukaryotes, flagella play important roles both in motility and as sensory organelles that monitor the extracellular environment. In the parasitic protozoan Leishmania mexicana, one glucose transporter isoform, LmxGT1, is targeted selectively to the flagellar membrane where it appears to play a role in glucose sensing. Trafficking of LmxGT1 to the flagellar membrane is dependent upon interaction with the KHARON1 protein that is located at the base of the flagellar axoneme. Remarkably, while Δkharon1 null mutants are viable as insect stage promastigotes, they are unable to survive as amastigotes inside host macrophages. Although Δkharon1 promastigotes enter macrophages and transform into amastigotes, these intracellular parasites are unable to execute cytokinesis and form multinucleate cells before dying. Notably, extracellular axenic amastigotes of Δkharon1 mutants replicate and divide normally, indicating a defect in the mutants that is only exhibited in the intra-macrophage environment. Although the flagella of Δkharon1 amastigotes adhere to the phagolysomal membrane of host macrophages, the morphology of the mutant flagella is often distorted. Additionally, these null mutants are completely avirulent following injection into BALB/c mice, underscoring the critical role of the KHARON1 protein for viability of intracellular amastigotes and disease in the animal model of leishmaniasis. PMID:26266938

  8. Transgenic, Fluorescent Leishmania mexicana Allow Direct Analysis of the Proteome of Intracellular Amastigotes*S⃞

    PubMed Central

    Paape, Daniel; Lippuner, Christoph; Schmid, Monika; Ackermann, Renate; Barrios-Llerena, Martin E.; Zimny-Arndt, Ursula; Brinkmann, Volker; Arndt, Benjamin; Pleissner, Klaus Peter; Jungblut, Peter R.; Aebischer, Toni

    2008-01-01

    Investigating the proteome of intracellular pathogens is often hampered by inadequate methodologies to purify the pathogen free of host cell material. This has also precluded direct proteome analysis of the intracellular, amastigote form of Leishmania spp., protozoan parasites that cause a spectrum of diseases that affect some 12 million patients worldwide. Here a method is presented that combines classic, isopycnic density centrifugation with fluorescent particle sorting for purification by exploiting transgenic, fluorescent parasites to allow direct proteome analysis of the purified organisms. By this approach the proteome of intracellular Leishmania mexicana amastigotes was compared with that of extracellular promastigotes that are transmitted by insect vectors. In total, 509 different proteins were identified by mass spectrometry and database search. This number corresponds to ∼6% of gene products predicted from the reference genome of Leishmania major. Intracellular amastigotes synthesized significantly more proteins with basic pI and showed a greater abundance of enzymes of fatty acid catabolism, which may reflect their living in acidic habitats and metabolic adaptation to nutrient availability, respectively. Bioinformatics analyses of the genes corresponding to the protein data sets produced clear evidence for skewed codon usage and translational bias in these organisms. Moreover analysis of the subset of genes whose products were more abundant in amastigotes revealed characteristic sequence motifs in 3′-untranslated regions that have been linked to translational control elements. This suggests that proteome data sets may be used to identify regulatory elements in mRNAs. Last but not least, at 6% coverage the proteome identified all vaccine antigens tested to date. Thus, the present data set provides a valuable resource for selection of candidate vaccine antigens. PMID:18474515

  9. Anti-inflammatory effects of phenolic compounds isolated from the flowers of Nymphaea mexicana Zucc.

    PubMed

    Hsu, Chin-Lin; Fang, Song-Chwan; Yen, Gow-Chin

    2013-08-01

    Nymphaea mexicana Zucc. is an aquatic plant species which belongs to the family Nymphaea and is commonly known as the yellow water lily. The aim of this work was to study the in vitro antiinflammatory effects of phenolic compounds isolated from the flowers of Nymphaea mexicana Zucc. Seven phenolic compounds including vanillic acid, 4-methoxy-3,5-dihydroxybenzoic acid, (2R,3R)-3,7-dihydroxyflavanone, naringenin (4), kaempferol 3-O-(3-O-acetyl-a-L-rhamnopyranoside), kaempferol 3-O-(2-O-acetyl-a-L-rhamnopyranoside), and quercetin 3-(30 0-acetylrhamnoside) (7) were isolated from the flowers of Nymphaea mexicana Zucc. These results revealed that compound 4 has the most prominent inhibitory effect on the LPS-stimulated nitric oxide (NO), monocyte chemotactic protein-1 (MCP-1), and tumor necrosis factor-alpha (TNF-a) production in RAW 264.7 macrophages. In addition, compound 4 also inhibited LPS-mediated induction of protein expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), and phospho-ERK in RAW 264.7 macrophages. Thus, compound 4 from the flowers of Nymphaea mexicana Zucc. may provide a potential therapeutic approach for inflammation-associated disorders. PMID:23727892

  10. "La Hermandad" and Chicanas Organizing: The Community Rhetoric of the "Comision Femenil Mexicana Nacional" Organization

    ERIC Educational Resources Information Center

    Leon, Kendall Marie

    2013-01-01

    To address the need for situated accounts of community rhetoric, this article examines the legacy of the first Chicana feminist organization, the "Comision Femenil Mexicana Nacional (CFMN)." The CFMN and their archival collection provide[d] Chicanas an education about how to interpret, be and act in the world. To invent a rhetorical…

  11. Counterstories of College Persistence by Undocumented Mexicana Students: Navigating Race, Class, Gender, and Legal Status

    ERIC Educational Resources Information Center

    Munoz, Susana Maria; Maldonado, Marta Maria

    2012-01-01

    This paper draws from four sets of four in-depth interviews and one subsequent focus group to examine how undocumented Mexicana students navigate identities and the meanings of race, gender, class, and legal status. We mobilize a critical race theory framework to center and explore the content of students' counterstories. While majoritarian…

  12. The california poppy (eschscholtzia mexicana) as a copper indicator plant - a new example

    USGS Publications Warehouse

    Chaffee, M.A.; Gale, C.W., III

    1976-01-01

    The abundance and distribution of the California poppy (Eschscholtzia mexicana) correlates closely with the copper-rich outcrop of a small porphyry-type deposit in Arizona. Chemical factors are probably more important than physical factors in determining why this species is sometimes found as a copper indicator plant. ?? 1976.

  13. Phosphoglycan repeat-deficient Leishmania mexicana parasites remain infectious to macrophages and mice.

    PubMed

    Ilg, T; Demar, M; Harbecke, D

    2001-02-16

    The human pathogen Leishmania synthesizes phosphoglycans (PGs) formed by variably modified phosphodisaccharide [6-Galbeta1-4Manalpha1-PO(4)] repeats and mannooligosaccharide phosphate [(Manalpha1-2)(0-5)Manalpha1-PO(4)] caps that occur lipid-bound on lipophosphoglycan, protein-bound on proteophosphoglycans, and as an unlinked form. PG repeat synthesis has been described as essential for survival and development of Leishmania throughout their life cycle, including for virulence to the mammalian host. In this study, this proposal was investigated in Leishmania mexicana using a spontaneous mutant that was fortuitously isolated from an infected mouse, and by generating a lmexlpg2 gene deletion mutant (Deltalmexlpg2), that lacks a Golgi GDP-Man transporter. The spontaneous mutant lacks PG repeats but synthesizes normal levels of mannooligosaccharide phosphate caps, whereas the Deltalmexlpg2 mutant is deficient in PG repeat synthesis and down-regulates cap expression. In contrast to expectations, both L. mexicana mutants not only retain their ability to bind to macrophages, but are also indistinguishable from wild type parasites with respect to colonization of and multiplication within host cells. Moreover, in mouse infection studies, the spontaneous L. mexicana repeat-deficient mutant and the Deltalmexlpg2 mutant showed no significant difference to a wild type strain with respect to the severity of disease caused by these parasites. Therefore, at least in Leishmania mexicana, PG repeat synthesis is not an absolute requirement for virulence. PMID:11071892

  14. Recurrence of Mexican long-tongued bats (Choeronycteris mexicana) at historical sites in Arizona and New Mexico

    USGS Publications Warehouse

    Cryan, P.M.; Bogan, M.A.

    2003-01-01

    The Mexican long-tongued bat (Choeronycteris mexicana) is 1 of 3 migratory, nectarivorous bats that seasonally occur in the extreme southwestern United States (US); the other 2 species are Leptonycteris curasoae and L. nivalis. Unlike the species of Leptonycteris, C. mexicana is not known to form large maternity colonies and is rarely encountered in groups of more than 12 individuals (Hoffmeister 1986). Possibly because of a propensity to form small roosting groups, the number of C. mexicana historically encountered is relatively low compared to other bat species. Although the range of C. mexicana extends from the southwestern United States into Honduras, less than 1500 individuals have been documented since its discovery in 1844 (Petryszyn and Cockrum In Press). Roosting and habitat needs of C. mexicana are poorly understood and it is unclear how such requirements might influence the apparent scarcity of these bats. Choeronycteris mexicana is known to roost in a variety of situations, typically in shallow caves or near the entrances of more extensive structures (Arroyo-Cabrales et al. 1987). Roost sites have been reported from various vegetation zones, including tropical deciduous forests at southern latitudes (Davis and Russell 1954), but roosts are frequently found in oak-conifer woodlands in the northern part of its range (Hoffmeister 1986). Mexican long-tongued bats are known to feed on nectar, pollen, or fruit of various flowering plants throughout their range (Gardner 1977). Although mutualistic relationships likely exist between C. mexicana and its food plants, very little is known about the role that this species plays as a pollinator or seed disperser of such plants. The identification and elucidation of mutualistic relationships are necessary steps toward effectively conserving ecosystems in the southwestern US (Allen-Wardell et al. 1998). Given the potential importance of C. mexicana as a pollinator and its apparent scarcity in the southwest US, current

  15. Molecular diagnosis of Leishmania mexicana in a cutaneous leishmaniasis case in Sinaloa, Mexico.

    PubMed

    Ochoa-Diaz, Yssete O; Lopez-Moreno, Carmina Y; Rendon-Maldonado, Jose G; Lopez-Moreno, Hector S

    2012-01-01

    Leishmaniasis has been considered endemic in Sinaloa, Mexico, since 1994. Despite that Leishmania mexicana is the main etiological agent of cutaneous leishmaniasis (CL) in other regions of Mexico, the species causing CL in patients from Sinaloa state has not been previously established, although Leishmania braziliensis has been found in the neighboring southern state, Nayarit. L. braziliensis is also associated with mucocutaneous leishmaniasis, which is a more complicated clinical variant. Due to the implications on individual and public health, the objective of this report was to identify the Leishmania species present in Sinaloa, Mexico. Using the first internal transcribed spacer (ITS-1) polymerase chain reaction-restriction fragment length polymorphism, we identified L. mexicana in a CL patient from Sinaloa and confirmed the extended distribution of this parasite in Mexico. PMID:21923263

  16. Past, Present and Future: Forty Years of Revista Mexicana de Astronomia y Astrofisica

    NASA Astrophysics Data System (ADS)

    Allen, Christine; Torres-Peimbert, Silvia

    2015-08-01

    We cast a retrospective view on 40 years of publishing the Revista Mexicana de Astronomía y Astrofísica, founded in 1974. The journal is peer-reviewed, has appeared regularly since its foundation, and continues to attract original research papers, mostly by Mexican and Latin American authors. We share some musings about the future of our journal, in view of recent developments in the scientific publishing field.

  17. Chemical composition and antifungal activity of essential oil of Chrysactinia mexicana gray.

    PubMed

    Cárdenas-Ortega, Norma C; Zavala-Sánchez, Miguel A; Aguirre-Rivera, J Rogelio; Pérez-González, Cuauhtémoc; Pérez-Gutiérrez, Salud

    2005-06-01

    The chemical composition of the essential oil of Chysactinia mexicana was analyzed by gas chromatography-mass spectrometry. Seventeen compounds were characterized; eucalyptol (41.3%), piperitone (37.7%), and linalyl acetate (9.1%) were found as the major components. The essential oil of leaves and piperitone completely inhibited Aspergillus flavus growth at relatively low concentrations (1.25 and 0.6 mg/mL, respectively). PMID:15913293

  18. Mg/Ca temperature calibration for the benthic foraminifers Bulimina inflata and Bulimina mexicana

    NASA Astrophysics Data System (ADS)

    Grunert, Patrick; Rosenthal, Yair; Jorissen, Frans; Holbourn, Ann

    2016-04-01

    Bulimina inflata Seguenza 1862 and Bulimina mexicana Cushman 1922 are cosmopolitan, shallow infaunal benthic foraminifers which are common in the fossil record throughout the Neogene and Quaternary. The closely related species share a similar costate shell morphology that differs in the presence or absence of an apical spine. In the present study, we evaluate the temperature dependency of Mg/Ca ratios of these species from an extensive set of core-top samples from the Atlantic and Pacific oceans. The results show no significant offset in Mg/Ca values between B. inflata, B. mexicana, and two other costate morphospecies when present in the same sample. The apparent lack of significant inter-specific/inter-morphotype differences amongst the analysed costate buliminds allows for the combined use of their data-sets for our core-top calibration. Over a bottom-water temperature range of 3-14°C, the Bulimina inflata/mexicana group shows a sensitivity of ˜0.12 mmol/mol/°C which is comparable to the epifaunal Cibicidoides pachyderma and higher than for the shallow infaunal Uvigerina spp., the most commonly used taxa in Mg/Ca-based palaeotemperature reconstruction. B. inflata and B. mexicana might thus be a valuable alternative in mesotrophic settings where many of the commonly used species are diminished or absent, and particularly useful in hypoxic settings where costate buliminds may dominate foraminiferal assemblages. This study was financially supported by the Max-Kade-Foundation and contributes to project P25831-N29 of the Austrian Science Fund (FWF).

  19. Characterization of a novel GDP-mannose:Serine-protein mannose-1-phosphotransferase from Leishmania mexicana.

    PubMed

    Moss, J M; Reid, G E; Mullin, K A; Zawadzki, J L; Simpson, R J; McConville, M J

    1999-03-01

    Protozoan parasites of the genus Leishmania secrete a number of glycoproteins and mucin-like proteoglycans that appear to be important parasite virulence factors. We have previously proposed that the polypeptide backbones of these molecules are extensively modified with a complex array of phosphoglycan chains that are linked to Ser/Thr-rich domains via a common Manalpha1-PO4-Ser linkage (Ilg, T., Overath, P., Ferguson, M. A. J., Rutherford, T., Campbell, D. G., and McConville, M. J. (1994) J. Biol. Chem. 269, 24073-24081). In this study, we show that Leishmania mexicana promastigotes contain a peptide-specific mannose-1-phosphotransferase (pep-MPT) activity that adds Manalpha1-P to serine residues in a range of defined peptides. The presence and location of the Manalpha1-PO4-Ser linkage in these peptides were determined by electrospray ionization mass spectrometry and chemical and enzymatic treatments. The pep-MPT activity was solubilized in non-ionic detergents, was dependent on Mn2+, utilized GDP-Man as the mannose donor, and was expressed in all developmental stages of the parasite. The pep-MPT activity was maximal against peptides containing Ser/Thr-rich domains of the endogenous acceptors and, based on competition assays with oligosaccharide acceptors, was distinct from other leishmanial MPTs involved in the initiation and elongation of lipid-linked phosphoglycan chains. In subcellular fractionation experiments, pep-MPT was resolved from the endoplasmic reticulum marker BiP, but had an overlapping distribution with the cis-Golgi marker Rab1. Although Man-PO4 residues in the mature secreted glycoproteins are extensively modified with mannose oligosaccharides and phosphoglycan chains, similar modifications were not added to peptide-linked Man-PO4 residues in the in vitro assays. Similarly, Man-PO4 residues on endogenous polypeptide acceptors were also poorly extended, although the elongating enzymes were still active, suggesting that the pep-MPT activity and

  20. Variation and Genetic Structure in Platanus mexicana (Platanaceae) along Riparian Altitudinal Gradient

    PubMed Central

    Galván-Hernández, Dulce M.; Lozada-García, J. Armando; Flores-Estévez, Norma; Galindo-González, Jorge; Vázquez-Torres, S. Mario

    2015-01-01

    Platanus mexicana is a dominant arboreal species of riparian ecosystems. These ecosystems are associated with altitudinal gradients that can generate genetic differences in the species, especially in the extremes of the distribution. However, studies on the altitudinal effect on genetic variation to riparian species are scarce. In Mexico, the population of P. mexicana along the Colipa River (Veracruz State) grows below its reported minimum altitude range, possibly the lowest where this tree grows. This suggests that altitude might be an important factor in population genetics differentiation. We examined the genetic variation and population structuring at four sites with different altitudes (70, 200, 600 and 1700 m a.s.l.) using ten inter-simple sequence repeats (ISSR) markers. The highest value for Shannon index and Nei’s gene diversity was obtained at 1700 m a.s.l. (He = 0.27, Ne = 1.47, I = 0.42) and polymorphism reached the top value at the middle altitude (% p = 88.57). Analysis of molecular variance (AMOVA) and STRUCTURE analysis indicated intrapopulation genetic differentiation. The arithmetic average (UPGMA) dendrogram identified 70 m a.s.l. as the most genetically distant site. The genetic structuring resulted from limited gene flow and genetic drift. This is the first report of genetic variation in populations of P. mexicana in Mexico. This research highlights its importance as a dominant species, and its ecological and evolutionary implications in altitudinal gradients of riparian ecosystems. PMID:25607732

  1. Transplacental Transmission of Cutaneous Leishmania mexicana Strain in BALB/c Mice

    PubMed Central

    Avila-García, Miroslava; Mancilla-Ramírez, Javier; Segura-Cervantes, Enrique; Farfan-Labonne, Blanca; Ramírez-Ramírez, Alicia; Galindo-Sevilla, Norma

    2013-01-01

    The vertical transmission of leishmaniasis has been reported in species that cause visceral leishmaniasis. However, this condition has scarcely been documented in species that cause cutaneous leishmaniasis. The aim of this study was to determine experimentally whether L. mexicana is transmitted vertically. A control group of BALB/c mice and a group infected with L. mexicana were mated, the gestation was monitored, and females were killed before delivery. Four resorptions (P = 0.023) and eight fetal deaths (P = 0.010) were observed in the infected female group; furthermore, the offspring body weight of the infected group was lower than the body weight of the healthy group (P = 0.009). DNA amplification by polymerase chain reaction (PCR) revealed that all placentas and maternal spleens as well as 39 of 110 fetal spleens obtained from the offspring of infected mothers tested positive for Leishmania. In conclusion, L. mexicana is transmitted transplacentally and causes fetal death, resorption, and reduction in offspring body weight. PMID:23798582

  2. Effect of neonatal injection with antibodies to Leishmania mexicana on its growth in adult infected mice.

    PubMed Central

    Gorczynski, R M

    1988-01-01

    Mice inoculated with monoclonal antibodies (MAb) directed to Leishmania mexicana antigens were not protected from growth of a subsequent challenge infection; this was the case even when those antibodies were capable of inhibiting parasite growth in vitro. However F(ab')2 fragments of one antibody (1E1) were protective in vivo. When neonatal mice were injected with MAb and subsequently infected as adults, the animals were more susceptible to parasite growth than uninjected controls. This increased susceptibility could be adoptively transferred with Lyt-1+ cells. Separate groups of animals were immunized with different MAb to L. mexicana, and parasite growth in these animals was studied. In no case was parasite growth altered, though these mice did produce specific antibodies directed against the immunizing MAb (anti-idiotypic antibodies). When neonatal mice were injected with these latter reagents, they were found to be more resistant to challenge infection than control animals. This resistance was associated with an enhanced ability of spleen cells from these mice to produce, on stimulation with parasite antigens in vitro, a factor rendering normal macrophages cytocidal for L. mexicana. PMID:2965682

  3. Cysteine Peptidase B Regulates Leishmania mexicana Virulence through the Modulation of GP63 Expression

    PubMed Central

    Casgrain, Pierre-André; Martel, Caroline; McMaster, W. Robert; Mottram, Jeremy C.; Olivier, Martin; Descoteaux, Albert

    2016-01-01

    Cysteine peptidases play a central role in the biology of Leishmania. In this work, we sought to further elucidate the mechanism(s) by which the cysteine peptidase CPB contributes to L. mexicana virulence and whether CPB participates in the formation of large communal parasitophorous vacuoles induced by these parasites. We initially examined the impact of L. mexicana infection on the trafficking of VAMP3 and VAMP8, two endocytic SNARE proteins associated with phagolysosome biogenesis and function. Using a CPB-deficient mutant, we found that both VAMP3 and VAMP8 were down-modulated in a CPB-dependent manner. We also discovered that expression of the virulence-associated GPI-anchored metalloprotease GP63 was inhibited in the absence of CPB. Expression of GP63 in the CPB-deficient mutant was sufficient to down-modulate VAMP3 and VAMP8. Similarly, episomal expression of GP63 enabled the CPB-deficient mutant to establish infection in macrophages, induce the formation of large communal parasitophorous vacuoles, and cause lesions in mice. These findings implicate CPB in the regulation of GP63 expression and provide evidence that both GP63 and CPB are key virulence factors in L. mexicana. PMID:27191844

  4. Antispasmodic Effects and Action Mechanism of Essential Oil of Chrysactinia mexicana A. Gray on Rabbit Ileum.

    PubMed

    Zavala-Mendoza, Daniel; Grasa, Laura; Zavala-Sánchez, Miguel Ángel; Pérez-Gutiérrez, Salud; Murillo, María Divina

    2016-01-01

    The Chrysactinia mexicana A. Gray (C. mexicana) plant is used in folk medicine to treat fever and rheumatism; it is used as a diuretic, antispasmodic; and it is used for its aphrodisiac properties. This study investigates the effects of the essential oil of C. mexicana (EOCM) on the contractility of rabbit ileum and the mechanisms of action involved. Muscle contractility studies in vitro in an organ bath to evaluate the response to EOCM were performed in the rabbit ileum. EOCM (1-100 µg·mL(-1)) reduced the amplitude and area under the curve of spontaneous contractions of the ileum. The contractions induced by carbachol 1 µM, potassium chloride (KCl) 60 mM or Bay K8644 1 µM were reduced by EOCM (30 µg·mL(-1)). Apamin 1 µM and charybdotoxin 0.01 µM decreased the inhibition induced by EOCM. The d-cAMP 1 µM decreased the inhibition induced by EOCM. l-NNA 10 µM, Rp-8-Br-PET-cGMPS 1 µM, d,l-propargylglycine 2 mM, or aminooxyacetic acid hemihydrochloride 2 mM did not modify the EOCM effect. In conclusion, EOCM induces an antispasmodic effect and could be used in the treatment of intestinal spasms or diarrhea processes. This effect would be mediated by Ca(2+), Ca(2+)-activated K⁺ channels and cAMP. PMID:27322223

  5. Leishmania mexicana Infection Induces IgG to Parasite Surface Glycoinositol Phospholipids that Can Induce IL-10 in Mice and Humans

    PubMed Central

    Buxbaum, Laurence U.

    2013-01-01

    Infection with the intracellular protozoan parasite Leishmania mexicana causes chronic disease in C57BL/6 mice, in which cutaneous lesions persist for many months with high parasite burdens (107–108 parasites). This chronic disease process requires host IL-10 and FcγRIII. When Leishmania amastigotes are released from cells, surface-bound IgG can induce IL-10 and suppress IL-12 production from macrophages. These changes decrease IFN-γ from T cells and nitric oxide production in infected cells, which are both required for Leishmania control. However, antibodies targets and the kinetics of antibody production are unknown. Several groups have been unsuccessful in identifying amastigote surface proteins that bind IgG. We now show that glycoinositol phospholipids (GIPLs) of L. mexicana are recognized by mouse IgG1 by 6 weeks of infection, with a rapid increase between 12 and 16 weeks, consistent with the timing of chronic disease in C57BL/6 mice vs. healing in FcγRIII-deficient mice. A single prominent spot on TLC is recognized by IgG, and the glycolipid is a glycosyl phosphatidylinositol containing a branched mannose structure. We show that the lipid structure of the GIPL (the sn-2 fatty acid) is required for antibody recognition. This GIPL is abundant in L. mexicana amastigotes, rare in stationary-phase promastigotes, and absent in L. major, consistent with a role for antibodies to GIPLs in chronic disease. A mouse monoclonal anti-GIPL IgG recognizes GIPLs on the parasite surface, and induces IL-10 from macrophages. The current work also extends this mouse analysis to humans, finding that L. mexicana-infected humans with localized and diffuse cutaneous leishmaniasis have antibodies that recognize GIPLs, can bind to the surface of amastigotes, and can induce IL-10 from human monocytes. Further characterization of the target glycolipids will have important implications for drug and vaccine development and will elucidate the poorly understood role of glycolipids in

  6. Anxiolytic-like effect of ethanolic extract of Argemone mexicana and its alkaloids in Wistar rats

    PubMed Central

    Arcos-Martínez, Aideé Itzel; Muñoz-Muñiz, Omar David; Domínguez-Ortiz, Miguel Ángel; Saavedra-Vélez, Margarita Virginia; Vázquez-Hernández, Maribel; Alcántara-López, María Gabriela

    2016-01-01

    Objective: Argemone mexicana is a Papaveracea plant; some reports have shown their antibacterial, anti-cancer, sedative and probably anti-anxiety properties. From their aerial parts, flavonoids and alkaloids have been isolated, which are intrinsically related to some actions on the central nervous system. The aim of this study was to evaluate the anxiolytic-like effects of the plant, using its ethanolic extract and alkaloid-enriched extract obtained from fresh leaves. Material and Methods: Phytochemical screening was carried out together with evaluation of antioxidant capacity and the enrichment of alkaloids present in the extract. Subsequently, 100 and 200 mg/kg doses of ethanolic extract and alkaloid-enriched extract (200 µg/kg) were intraperitoneally administered to female Wistar rats, which were exposed to elevated plus maze (EPM) test. Picrotoxin (1 mg/kg), a non-competitive gamma-aminobutyric acid A (GABAA) chloride channel antagonist, was used in experimental procedures to evaluate if this receptor is involved in the anxiolytic-like effects of A. mexicana. To discard motor effects associated with the treatments, the rats were evaluated by the locomotor activity test. Results: Only the ethanolic extract at 200 mg/kg and alkaloid-enriched extract (200 µg/kg) produced anxiolytic-like effects similarly to diazepam 2 mg/kg on EPM test, without affecting locomotor activity. Meanwhile, the administration of picrotoxin blocked anti-anxiety effect of alkaloid-enriched extract of the plant. Conclusion: These results showed that A. mexicana is a potential anxiolytic agent and we suggest that this effect is mediated by the GABAA receptor. These effects are related to the presence of alkaloids. PMID:27516989

  7. Isolation of cDNA from Jacaratia mexicana encoding a mexicain-like cysteine protease gene.

    PubMed

    Ramos-Martínez, Erick M; Herrera-Ramírez, Alejandra C; Badillo-Corona, Jesús Agustín; Garibay-Orijel, Claudio; González-Rábade, Nuria; Oliver-Salvador, María Del Carmen

    2012-07-01

    Cysteine proteases (CPs) from the C1 family, which are similar to papain, can be found in animals and plants, as well as some viruses and prokaryotes. These enzymes have diverse physiological functions and are thus very attractive for science and industry. Jacaratia mexicana, a member of the Caricaceae plant family, contains several CPs, the principal being mexicain, found to favorably compete against papain for many industrial applications due to its high stability and specific activity. In this study, leaves of J. mexicana were used to isolate a CP-coding gene, similar to those that code for mexicain and chymomexicain. By using rapid amplification of cDNA ends (RACE) as well as oligonucleotide design from papain-like conserved amino acids (aa), a sequence of 1404 bp consisting of a 5' terminal untranslated region (UTR) of 153 bp, a 3' terminal UTR of 131 bp, with a polyadenylation (poly(A)) signal sequence and a poly(A) tail, and an open reading frame (ORF) of 1046 bp, was obtained by overlapping three partial sequences. Two full-length cDNA sequences that encode for mexicain-like proteases were cloned from mRNA (JmCP4 and JmCP5). JmCP4 is predicted to have an ORF of 1044 bp, which codifies for polypeptides that have a 26 aa signal peptide region, a 108 aa propeptide region and a mature enzyme of 214 aa. A 969 bp fragment (JmCP5) encodes for a partial sequence of a CP gene, without the signal peptide region but with a full-length propeptide region. The sequence analysis showed that this protease presented a high similarity to other plant CPs from J. mexicana, Vasconcellea cundinamarcensis, Vasconcellea stipulata, and Carica papaya, among others, mainly at the conserved catalytic site. Obtaining the sequence of this CP gene from J. mexicana provides an alternative for production in a standard system and could be an initial step towards the commercialization of this enzyme. PMID:22543019

  8. Differences in Lsh gene control over systemic Leishmania major and Leishmania donovani or Leishmania mexicana mexicana infections are caused by differential targeting to infiltrating and resident liver macrophage populations.

    PubMed Central

    Davies, E V; Singleton, A M; Blackwell, J M

    1988-01-01

    Earlier studies had shown that the viscerotropic NIH 173 strain of cutaneous Leishmania major fails to come under Lsh gene control. Visceral Leishmania donovani LV9 and another viscerotropic cutaneous strain, Leishmania mexicana mexicana LV4, are controlled by Lsh. The results of double-infection experiments presented here show that expression of Lsh resistance against L. mexicana mexicana was enhanced in the presence of L. donovani, whereas L. major still failed to come under Lsh gene control, even in the presence of L. donovani. Prior irradiation (850 rads) of mice showed that in the absence of infiltrating monocytes, Lsh did exert some influence over L. major. The presence of a higher infiltrate of fresh monocytes after L. major infection was confirmed in liver macrophage populations isolated from mice after infection in vivo and in liver cryosections immunostained with monoclonal antibody M1/70 directed against the type 3 complement receptor CR3. The results support the hypothesis that Lsh is expressed maximally in the resident tissue macrophages and poorly in the immature macrophages preferentially infected by L. major amastigotes. Images PMID:3356462

  9. Identification of Semicarbazones, Thiosemicarbazones and Triazine Nitriles as Inhibitors of Leishmania mexicana Cysteine Protease CPB

    PubMed Central

    Schröder, Jörg; Noack, Sandra; Marhöfer, Richard J.; Mottram, Jeremy C.; Coombs, Graham H.; Selzer, Paul M.

    2013-01-01

    Cysteine proteases of the papain superfamily are present in nearly all eukaryotes. They play pivotal roles in the biology of parasites and inhibition of cysteine proteases is emerging as an important strategy to combat parasitic diseases such as sleeping sickness, Chagas’ disease and leishmaniasis. Homology modeling of the mature Leishmania mexicana cysteine protease CPB2.8 suggested that it differs significantly from bovine cathepsin B and thus could be a good drug target. High throughput screening of a compound library against this enzyme and bovine cathepsin B in a counter assay identified four novel inhibitors, containing the warhead-types semicarbazone, thiosemicarbazone and triazine nitrile, that can be used as leads for antiparasite drug design. Covalent docking experiments confirmed the SARs of these lead compounds in an effort to understand the structural elements required for specific inhibition of CPB2.8. This study has provided starting points for the design of selective and highly potent inhibitors of L. mexicana cysteine protease CPB that may also have useful efficacy against other important cysteine proteases. PMID:24146999

  10. Vegetation patches improve the establishment of Salvia mexicana seedlings by modifying microclimatic conditions

    NASA Astrophysics Data System (ADS)

    Mendoza-Hernández, Pedro E.; Rosete-Rodríguez, Alejandra; Sánchez-Coronado, María E.; Orozco, Susana; Pedrero-López, Luis; Méndez, Ignacio; Orozco-Segovia, Alma

    2014-07-01

    Human disturbance has disrupted the dynamics of plant communities. To restore these dynamics, we could take advantage of the microclimatic conditions generated by remaining patches of vegetation and plastic mulch. These microclimatic conditions might have great importance in restoring disturbed lava fields located south of Mexico City, where the rock is exposed and the soil is shallow. We evaluated the effects of both the shade projected by vegetation patches and plastic mulch on the mean monthly soil surface temperature ( T ss) and photosynthetic photon flux density (PPFD) and on the survival and growth of Salvia mexicana throughout the year. This species was used as a phytometer of microsite quality. Shade reduced the T ss to a greater extent than mulch did. Both survival and growth were enhanced by shade and mulch, and the PPFD was related with seedling growth. During the dry season, plant biomass was lost, and there was a negative effect of PPFD on plant growth. At micro-meteorological scales, the use of shade projected by patches of vegetation and mulch significantly reduced the mortality of S. mexicana and enhanced its growth. Survival and growth of this plant depended on the environmental quality of microsites on a small scale, which was determined by the environmental heterogeneity of the patches and the landscape. For plant restoration, microsite quality must be evaluated on small scales, but on a large scale it may be enough to take advantage of landscape shade dynamics and the use of mulch to increase plant survival and growth.

  11. Vegetation patches improve the establishment of Salvia mexicana seedlings by modifying microclimatic conditions

    NASA Astrophysics Data System (ADS)

    Mendoza-Hernández, Pedro E.; Rosete-Rodríguez, Alejandra; Sánchez-Coronado, María E.; Orozco, Susana; Pedrero-López, Luis; Méndez, Ignacio; Orozco-Segovia, Alma

    2013-04-01

    Human disturbance has disrupted the dynamics of plant communities. To restore these dynamics, we could take advantage of the microclimatic conditions generated by remaining patches of vegetation and plastic mulch. These microclimatic conditions might have great importance in restoring disturbed lava fields located south of Mexico City, where the rock is exposed and the soil is shallow. We evaluated the effects of both the shade projected by vegetation patches and plastic mulch on the mean monthly soil surface temperature (T ss) and photosynthetic photon flux density (PPFD) and on the survival and growth of Salvia mexicana throughout the year. This species was used as a phytometer of microsite quality. Shade reduced the T ss to a greater extent than mulch did. Both survival and growth were enhanced by shade and mulch, and the PPFD was related with seedling growth. During the dry season, plant biomass was lost, and there was a negative effect of PPFD on plant growth. At micro-meteorological scales, the use of shade projected by patches of vegetation and mulch significantly reduced the mortality of S. mexicana and enhanced its growth. Survival and growth of this plant depended on the environmental quality of microsites on a small scale, which was determined by the environmental heterogeneity of the patches and the landscape. For plant restoration, microsite quality must be evaluated on small scales, but on a large scale it may be enough to take advantage of landscape shade dynamics and the use of mulch to increase plant survival and growth.

  12. Experience, but not distance, influences the recruitment precision in the stingless bee Scaptotrigona mexicana

    NASA Astrophysics Data System (ADS)

    Sánchez, Daniel; Kraus, F. Bernhard; Hernández, Manuel De Jesús; Vandame, Rémy

    2007-07-01

    Recruitment precision, i.e. the proportion of recruits that reach an advertised food source, is a crucial adaptation of social bees to their environment. Studies with honeybees showed that recruitment precision is not a fixed feature, but it may be enhanced by factors like experience and distance. However, little is known regarding the recruitment precision of stingless bees. Hence, in this study, we examined the effects of experience and spatial distance on the precision of the food communication system of the stingless bee Scaptotrigona mexicana. We conducted the experiments by training bees to a three-dimensional artificial patch at several distances from the colony. We recorded the choices of individual recruited foragers, either being newcomers (foragers without experience with the advertised food source) or experienced (foragers that had previously visited the feeder). We found that the average precision of newcomers (95.6 ± 2.61%) was significantly higher than that of experienced bees (80.2 ± 1.12%). While this might seem counter-intuitive on first sight, this “loss” of precision can be explained by the tendency of experienced recruits to explore nearby areas to find new rewarding food sources after they had initially learned the exact location of the food source. Increasing the distance from the colony had no significant effect on the precision of the foraging bees. Thus, our data show that experience, but not the distance of the food source, affected the patch precision of S. mexicana foragers.

  13. Parasitophorous vacuoles of Leishmania mexicana acquire macromolecules from the host cell cytosol via two independent routes.

    PubMed

    Schaible, U E; Schlesinger, P H; Steinberg, T H; Mangel, W F; Kobayashi, T; Russell, D G

    1999-03-01

    The intracellular parasite Leishmania survives and proliferates in host macrophages. In this study we show that parasitophorous vacuoles of L. mexicana gain access to cytosolic material via two different routes. (1) Small anionic molecules such as Lucifer Yellow are rapidly transported into the vacuoles by an active transport mechanism that is sensitive to inhibitors of the host cell's organic anion transporter. (2) Larger molecules such as fluorescent dextrans introduced into the host cell cytosol are also delivered to parasitophorous vacuoles. This transport is slower and sensitive to modulators of autophagy. Infected macrophages were examined by two novel assays to visualize and quantify this process. Immunoelectron microscopy of cells loaded with digoxigenin-dextran revealed label in multivesicular endosomes, which appeared to fuse with parasitophorous vacuoles. The inner membranes of the multivesicular vesicles label strongly with antibodies against lysobisphosphatidic acid, suggesting that they represent a point of confluence between the endosomal and autophagosomal pathways. Although the rate of autophagous transfer was comparable in infected and uninfected cells, infected cells retained hydrolyzed cysteine proteinase substrate to a greater degree. These data suggest that L. mexicana-containing vacuoles have access to potential nutrients in the host cell cytosol via at least two independent mechanisms. PMID:9973603

  14. The histopathology of cutaneous leishmaniasis due to Leishmania (Leishmania) mexicana in the Yucatan peninsula, Mexico.

    PubMed

    Andrade-Narvaez, Fernando J; Medina-Peralta, Salvador; Vargas-Gonzalez, Alberto; Canto-Lara, Silvia B; Estrada-Parra, Sergio

    2005-01-01

    Localized Cutaneous Leishmaniasis (LCL) known as "chiclero's ulcer" in southeast Mexico, was described by SEIDELIN in 1912. Since then the sylvatic region of the Yucatan peninsula has been documented as an endemic focus of LCL. This study of 73 biopsies from parasitological confirmed lesions of LCL cases of Leishmania (Leishmania) mexicana infection was undertaken: 1) to examine host response at tissue level; and 2) to relate manifestations of this response to some characteristics of clinical presentation. Based on Magalhães' classification we found that the most common pattern in our LCL cases caused by L. (L.) mexicana was predominantly characterized by the presence of unorganized granuloma without necrosis, (43.8%). Another important finding to be highlighted is the fact that in 50/73 (68.5%) parasite identification was positive. There was direct relation between the size of the lesion and time of evolution (rs = 0.3079, p = 0.03), and inverse correlation between size of the lesion and abundance of amastigotes (rs = -0.2467, p = 0.03). In view of the complexity of clinical and histopathological findings, cell-mediated immune response of the disease related to clinical and histopathological features, as so genetic background should be studied. PMID:16138198

  15. The role of phosphomannose isomerase in Leishmania mexicana glycoconjugate synthesis and virulence.

    PubMed

    Garami, A; Ilg, T

    2001-03-01

    Phosphomannose isomerase (PMI) catalyzes the reversible interconversion of fructose 6-phosphate and mannose 6-phosphate, which is the first step in the biosynthesis of activated mannose donors required for the biosynthesis of various glycoconjugates. Leishmania species synthesize copious amounts of mannose-containing glycolipids and glycoproteins, which are involved in virulence of these parasitic protozoa. To investigate the role of PMI for parasite glycoconjugate synthesis, we have cloned the PMI gene (lmexpmi) from Leishmania mexicana, generated gene deletion mutants (Delta lmexpmi), and analyzed their phenotype. Delta lmexpmi mutants lack completely the high PMI activity found in wild type parasites, but are, in contrast to fungi, able to grow in media deficient for free mannose. The mutants are unable to synthesize phosphoglycan repeats [-6-Gal beta 1-4Man alpha 1-PO(4)-] and mannose-containing glycoinositolphospholipids, and the surface expression of the glycosylphosphatidylinositol-anchored dominant surface glycoprotein leishmanolysin is strongly decreased, unless the parasite growth medium is supplemented with mannose. The Delta lmexpmi mutant is attenuated in infections of macrophages in vitro and of mice, suggesting that PMI may be a target for anti-Leishmania drug development. L. mexicana Delta lmexpmi provides the first conditional mannose-controlled system for parasite glycoconjugate assembly with potential applications for the investigation of their biosynthesis, intracellular sorting, and function. PMID:11084042

  16. Aqueous and Methanolic Extracts of Caulerpa mexicana Suppress Cell Migration and Ear Edema Induced by Inflammatory Agents

    PubMed Central

    Bitencourt, Mariana Angelica Oliveira; Dantas, Gracielle Rodrigues; Lira, Daysianne Pereira; Barbosa-Filho, Jose Maria; de Miranda, George Emmanuel Cavalcanti; de Oliveira Santos, Barbara Viviana; Souto, Janeusa Trindade

    2011-01-01

    The regulation of the inflammatory response is essential to maintaining homeostasis. Several studies have investigated new drugs that may contribute to avoiding or minimizing excessive inflammatory process. The aim of this study was to evaluate the effect of extracts of green algae Caulerpa mexicana on models inflammation. In mice, the inflammatory peritonitis model is induced by zymosan. Previous treatment of mice with aqueous and methanolic extracts of C. mexicana was able to suppress the cell migration to the peritoneal cavity, in a time-dependent but not in a dose-dependent manner. The treatment of mice with C. mexicana extracts also decreased the xylene-induced ear edema, exerting strong inhibitory leukocyte migration elicited by zymosan into the air pouch. We concluded that administration of the extracts resulted in a reduction of cell migration to different sites as well as a decrease in edema formation induced by chemical irritants. This study demonstrates for the first time the anti-inflammatory effect of aqueous and methanolic extracts from the green marine algae Caulerpa mexicana. PMID:21892348

  17. Revista Mexicana de Astronomía y Astrofísica, a real option for astronomical publication

    NASA Astrophysics Data System (ADS)

    Torres-Peimbert, S.; Allen, C.

    2011-10-01

    We present statistical data about the Revista Mexicana de Astronomía y Astrofísica. We consider that this journal is well positioned in the international astronomical literature. Similarly we present information about the Serie de Conferencias, which also has a wide level of acceptance by the astronomical community.

  18. Disease Severity in Patients Infected with Leishmania mexicana Relates to IL-1β

    PubMed Central

    Fernández-Figueroa, Edith A.; Rangel-Escareño, Claudia; Espinosa-Mateos, Valeria; Carrillo-Sánchez, Karol; Salaiza-Suazo, Norma; Carrada-Figueroa, Georgina; March-Mifsut, Santiago; Becker, Ingeborg

    2012-01-01

    Leishmania mexicana can cause both localized (LCL) and diffuse (DCL) cutaneous leishmaniasis, yet little is known about factors regulating disease severity in these patients. We analyzed if the disease was associated with single nucleotide polymorphisms (SNPs) in IL-1β (−511), CXCL8 (−251) and/or the inhibitor IL-1RA (+2018) in 58 Mexican mestizo patients with LCL, 6 with DCL and 123 control cases. Additionally, we analyzed the in vitro production of IL-1β by monocytes, the expression of this cytokine in sera of these patients, as well as the tissue distribution of IL-1β and the number of parasites in lesions of LCL and DCL patients. Our results show a significant difference in the distribution of IL-1β (−511 C/T) genotypes between patients and controls (heterozygous OR), with respect to the reference group CC, which was estimated with a value of 3.23, 95% CI = (1.2, 8.7) and p-value = 0.0167), indicating that IL-1β (−511 C/T) represents a variable influencing the risk to develop the disease in patients infected with Leishmania mexicana. Additionally, an increased in vitro production of IL-1β by monocytes and an increased serum expression of the cytokine correlated with the severity of the disease, since it was significantly higher in DCL patients heavily infected with Leishmania mexicana. The distribution of IL-1β in lesions also varied according to the number of parasites harbored in the tissues: in heavily infected LCL patients and in all DCL patients, the cytokine was scattered diffusely throughout the lesion. In contrast, in LCL patients with lower numbers of parasites in the lesions, IL-1β was confined to the cells. These data suggest that IL-1β possibly is a key player determining the severity of the disease in DCL patients. The analysis of polymorphisms in CXCL8 and IL-1RA showed no differences between patients with different disease severities or between patients and controls. PMID:22629474

  19. α-Ketoheterocycles as inhibitors of Leishmania mexicana cysteine protease CPB

    PubMed Central

    Steert, Koen; Berg, Maya; Mottram, Jeremy C.; Westrop, Gareth D.; Coombs, Graham H.; Cos, Paul; Maes, Louis; Joossens, Jurgen; Van der Veken, Pieter; Haemers, Achiel; Augustyns, Koen

    2011-01-01

    Cysteine proteases of the papain superfamily are present in nearly all eukaryotes and also play pivotal roles in the biology of parasites. Inhibition of cysteine proteases is emerging as an important strategy to combat parasitic diseases such as sleeping sickness, Chagas’ disease and leishmaniasis. Inspired by the in vivo antiparasitic activity of the vinyl sulfone based cysteine protease inhibitors (CPIs), a series of α-ketoheterocycles 1-15 has been developed as reversible inhibitors of a recombinant L. mexicana cysteine protease CPB2.8. The isoxazoles 1-3 and especially the oxadiazole 15 are potent reversible inhibitors of CPB2.8, however, in vitro whole-organism screening against a panel of protozoan parasites did not fully correlate with the observed inhibition of the cysteine protease. PMID:20799311

  20. Peltomexicanin, a Peltogynoid Quinone Methide from Peltogyne Mexicana Martínez Purple Heartwood.

    PubMed

    Gutiérrez-Macías, Paulina; Peralta-Cruz, Javier; Borja-de-la-Rosa, Amparo; Barragán-Huerta, Blanca E

    2016-01-01

    Peltomexicanin (7,10-dihydroxy-6,12-dioxa-5H-tetraphen-3-one) is a new peltogynoid quinone methide isolated from Palo Morado (Peltogyne mexicana Martínez) heartwood by column chromatography. Its chemical structure was elucidated by IR, NMR (¹H, (13)C), 2D NMR experiments (COSY, NOESY, HMQC, and HSQC), ESI-MS, and UV-Vis spectroscopic analysis. According to HPLC quantification, this compound is the main pigment and accounts for 1.21% of Palo Morado heartwood material. The antioxidant activity of peltomexicanin and dried methanolic extract (DEx) of purple heartwood was evaluated using the radical of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assay, and the corresponding values expressed as Trolox equivalents (µmol TE/mg sample) were 4.25 and 4.57, respectively. PMID:26861267

  1. Tourism values for Mexican free-tailed bat (Tadarida brasiliensis mexicana) viewing

    USGS Publications Warehouse

    Bagstad, Kenneth J.; Widerholdt, Ruscena

    2013-01-01

    Migratory species provide diverse ecosystem services to people, but these values have seldom been estimated rangewide for a single species. In this article, we summarize visitation and consumer surplus for recreational visitors to viewing sites for the Mexican free-tailed bat (Tadarida brasiliensis mexicana) throughout the Southwestern United States. Public bat viewing opportunities are available at 17 of 25 major roosts across six states; on an annual basis, we estimate that over 242,000 visitors view bats, gaining over $6.5 million in consumer surplus. A better understanding of spatial mismatches between the areas where bats provide value to people and areas most critical for maintaining migratory populations can better inform conservation planning, including economic incentive systems for conservation.

  2. Coxiella burnetii and Leishmania mexicana residing within similar parasitophorous vacuoles elicit disparate host responses

    PubMed Central

    Millar, Jess A.; Valdés, Raquel; Kacharia, Fenil R.; Landfear, Scott M.; Cambronne, Eric D.; Raghavan, Rahul

    2015-01-01

    Coxiella burnetii is a bacterium that thrives in an acidic parasitophorous vacuole (PV) derived from lysosomes. Leishmania mexicana, a eukaryote, has also independently evolved to live in a morphologically similar PV. As Coxiella and Leishmania are highly divergent organisms that cause different diseases, we reasoned that their respective infections would likely elicit distinct host responses despite producing phenotypically similar parasite-containing vacuoles. The objective of this study was to investigate, at the molecular level, the macrophage response to each pathogen. Infection of THP-1 (human monocyte/macrophage) cells with Coxiella and Leishmania elicited disparate host responses. At 5 days post-infection, when compared to uninfected cells, 1057 genes were differentially expressed (746 genes up-regulated and 311 genes down-regulated) in C. burnetii infected cells, whereas 698 genes (534 genes up-regulated and 164 genes down-regulated) were differentially expressed in L. mexicana infected cells. Interestingly, of the 1755 differentially expressed genes identified in this study, only 126 genes (~7%) are common to both infections. We also discovered that 1090 genes produced mRNA isoforms at significantly different levels under the two infection conditions, suggesting that alternate proteins encoded by the same gene might have important roles in host response to each infection. Additionally, we detected 257 micro RNAs (miRNAs) that were expressed in THP-1 cells, and identified miRNAs that were specifically expressed during Coxiella or Leishmania infections. Collectively, this study identified host mRNAs and miRNAs that were influenced by Coxiella and/or Leishmania infections, and our data indicate that although their PVs are morphologically similar, Coxiella and Leishmania have evolved different strategies that perturb distinct host processes to create and thrive within their respective intracellular niches. PMID:26300862

  3. Reverse pharmacology for developing an anti-malarial phytomedicine. The example of Argemone mexicana

    PubMed Central

    Simoes-Pires, Claudia; Hostettmann, Kurt; Haouala, Amina; Cuendet, Muriel; Falquet, Jacques; Graz, Bertrand; Christen, Philippe

    2014-01-01

    Classical pharmacology has been the basis for the discovery of new chemical entities with therapeutic effects for decades. In natural product research, compounds are generally tested in vivo only after full in vitro characterization. However drug screening using this methodology is expensive, time-consuming and very often inefficient. Reverse pharmacology, also called bedside-to-bench, is a research approach based on the traditional knowledge and relates to reversing the classical laboratory to clinic pathway to a clinic to laboratory practice. It is a trans-disciplinary approach focused on traditional knowledge, experimental observations and clinical experiences. This paper is an overview of the reverse pharmacology approach applied to the decoction of Argemone mexicana, used as an antimalarial traditional medicine in Mali. A. mexicana appeared as the most effective traditional medicine for the treatment of uncomplicated falciparum malaria in Mali, and the clinical efficacy of the decoction was comparable to artesunate–amodiaquine as previously published. Four stages of the reverse pharmacology process will be described here with a special emphasis on the results for stage 4. Briefly, allocryptopine, protopine and berberine were isolated through bioguided fractionation, and had their identity confirmed by spectroscopic analysis. The three alkaloids showed antiparasitic activity in vitro, of which allocryptopine and protopine were selective towards Plasmodiumfalciparum. Furthermore, the amount of the three active alkaloids in the decoction was determined by quantitative NMR, and preliminary in vivo assays were conducted. On the basis of these results, the reverse pharmacology approach is discussed and further pharmacokinetic studies appear to be necessary in order to determine whether these alkaloids can be considered as phytochemical markers for quality control and standardization of an improved traditional medicine made with this plant. PMID:25516845

  4. [Infection of skin fibroblasts in animals with different levels of sensitivity to Leishmania infantum and Leishmania mexicana (Kinetoplastida: Trypanosomatidae)].

    PubMed

    Minero, Miguel Angel; Chinchilla, Misael; Guerrero, Olga Marta; Castro, Alfredo

    2004-03-01

    Infection and multiplication of Leishmania infantum and L. mexicana inside of skin fibroblasts from hamsters, mice and rats was achieved. This process was demonstrated either by counting parasites inside the stained cells or by electronic microscopy studies. In addition multiplication rate differences in the cells from these rodent species were determined, for L. infantum as well as for L. mexicana. Parasite development in hamsters and mice fibroblasts was evident but there was not multiplication in rat cells showing that apparently they are refractory to Leishmania infection. These results suggest that the parasite affinity for each animal, as well as any intracellular environment resistance, could involve genetic factors in the parasite multiplication. On the other hand, presence of amastigote multiplication inside of parasitophorus vacuole, showed by electronic microscopy images, probes a true parasite transformation. Therefore it is suggested that fibroblasts could work as host cells for parasite survival and permanency in the infected animals. PMID:17357424

  5. Effect of Argemone mexicana (L.) against lithium-pilocarpine induced status epilepticus and oxidative stress in Wistar rats.

    PubMed

    Asuntha, G; Raju, Y Prasanna; Sundaresan, C R; Rasheed, Arun; Chowdary, V Harini; Vandana, K R; Babu, K Satish; Prasad, K V S R G

    2015-01-01

    Argemone mexicana (L.) has a role in the treatment of epileptic disorders in Indian traditional system of medicine. We studied its effect on induced status epilepticus (SE) and oxidative stress in rats. SE was induced in male albino rats by administration of pilocarpine (30 mg/kg, ip) 24 h after injection of lithium chloride (3 mEq/kg, ip). Different doses of the ethanol extract of A. mexicana were administered orally 1 h before the injection of pilocarpine. The severity of SE was observed and recorded every 15 min for 90 min and thereafter at every 30 min for another 90 min, using the Racine scoring system. In vivo lipid peroxidation of rat brain tissue was measured utilizing thiobarbiturate-reactive substances. Both in vitro free radical nitric oxide and 2,2-diphenyl-1-picryl hydrazyl scavenging activities of the extract were also determined. The SE severity was significantly reduced following oral administration of the extract at 250, 500 and 1000 mg/kg doses. None of the animals from groups 3 to 5 (with A. mexicana extract) have exhibited forelimb clonus of stage 4 seizure. The extract also exhibited both in vivo and in vitro antioxidant activities. PMID:25675709

  6. Glycosylation defects and virulence phenotypes of Leishmania mexicana phosphomannomutase and dolicholphosphate-mannose synthase gene deletion mutants.

    PubMed

    Garami, A; Mehlert, A; Ilg, T

    2001-12-01

    Leishmania parasites synthesize an abundance of mannose (Man)-containing glycoconjugates thought to be essential for virulence to the mammalian host and for viability. These glycoconjugates include lipophosphoglycan (LPG), proteophosphoglycans (PPGs), glycosylphosphatidylinositol (GPI)-anchored proteins, glycoinositolphospholipids (GIPLs), and N-glycans. A prerequisite for their biosynthesis is an ample supply of the Man donors GDP-Man and dolicholphosphate-Man. We have cloned from Leishmania mexicana the gene encoding the enzyme phosphomannomutase (PMM) and the previously described dolicholphosphate-Man synthase gene (DPMS) that are involved in Man activation. Surprisingly, gene deletion experiments resulted in viable parasite lines lacking the respective open reading frames (DeltaPMM and DeltaDPMS), a result against expectation and in contrast to the lethal phenotype observed in gene deletion experiments with fungi. L. mexicana DeltaDPMS exhibits a selective defect in LPG, protein GPI anchor, and GIPL biosynthesis, but despite the absence of these structures, which have been implicated in parasite virulence and viability, the mutant remains infectious to macrophages and mice. By contrast, L. mexicana DeltaPMM are largely devoid of all known Man-containing glycoconjugates and are unable to establish an infection in mouse macrophages or the living animal. Our results define Man activation leading to GDP-Man as a virulence pathway in Leishmania. PMID:11689705

  7. Complete set of mitochondrial pan-edited mRNAs in Leishmania mexicana amazonensis LV78

    PubMed Central

    Maslov, Dmitri A.

    2010-01-01

    Editing of mRNA transcribed from the mitochondrial cryptogenes ND8 (G1), ND9 (G2), G3, G4, ND3 (G5), RPS12 (G6) was investigated in Leishmania mexicana amazonensis, strain LV78, by amplification of the cDNA, cloning and sequencing. For each of these genes, extensively and partially edited transcripts were found to be relatively abundant compared to the respective pre-edited molecules. Moreover, the editing patterns observed in a majority of transcripts of each gene were consistent among themselves which allowed for inferring consensus editing sequences. The open reading frames contained in the consensus sequences were predicted to encode polypeptides that were highly similar to their counterparts in other species of Trypanosomatidae. Several kinetoplast DNA minicircles from this species available in the public domain were found to contain genes for guide RNAs which mediate editing of some of the mRNAs. The results indicate that the investigated strain of L. m. amazonensis has preserved its full editing capacity in spite of the long-term maintenance in culture. This property differs drastically from the other Leishmania species which lost some or all of the G1–G5 mRNA editing ability in culture. PMID:20546801

  8. Fatal anemia and dermatitis in captive agoutis (Dasyprocta mexicana) infested with Echidnophaga fleas.

    PubMed

    Cucchi-Stefanoni, Karina; Juan-Sallés, Carles; Parás, Alberto; Garner, Michael M

    2008-08-17

    Two captive agoutis (Dasyprocta mexicana) died of anemia with centrilobular hepatocellular necrosis (2/2), severe flea ectoparasitism (2/2), and cardiomegaly attributed to anemia (1/2). Other agoutis were similarly parasitized and one had anemia. Fleas were manually removed and all agoutis treated topically with propoxur and selamectin and moved to another enclosure. No additional cases of fatal anemia were seen. Cutaneous lesions suggestive of hypersensitivity were observed in three additional agoutis with dorsal alopecia (3/3), a penetrating wound associated with pruritus and self-mutilation in the flank (2/3), flea ectoparasitism at the time of morphologic diagnosis (1/3), and hyperplastic perivascular dermatitis (3/3). One of these died of bacterial infection of the wound. Similar but milder skin disease was seen in 3 out of over 30 maras (Dolichotis patagonum) housed in the same exhibit. Fleas collected from all the fatal agouti cases and maras were classified in the genus Echidnophaga based on the angular front margin of head, contracted thorax, absence of genal and pronotal combs, and the fact that fleas did not jump. These findings suggest that flea ectoparasitism may be an important cause of morbidity and mortality in captive rodents. PMID:18556127

  9. Microhabitat use, population densities, and size distributions of sulfur cave-dwelling Poecilia mexicana

    PubMed Central

    Bierbach, David; Riesch, Rüdiger; Schießl, Angela; Wigh, Adriana; Arias-Rodriguez, Lenin; Indy, Jeane Rimber; Klaus, Sebastian; Zimmer, Claudia; Plath, Martin

    2014-01-01

    The Cueva del Azufre in Tabasco, Mexico, is a nutrient-rich cave and its inhabitants need to cope with high levels of dissolved hydrogen sulfide and extreme hypoxia. One of the successful colonizers of this cave is the poeciliid fish Poecilia mexicana, which has received considerable attention as a model organism to examine evolutionary adaptations to extreme environmental conditions. Nonetheless, basic ecological data on the endemic cave molly population are still missing; here we aim to provide data on population densities, size class compositions and use of different microhabitats. We found high overall densities in the cave and highest densities at the middle part of the cave with more than 200 individuals per square meter. These sites have lower H2S concentrations compared to the inner parts where most large sulfide sources are located, but they are annually exposed to a religious harvesting ceremony of local Zoque people called La Pesca. We found a marked shift in size/age compositions towards an overabundance of smaller, juvenile fish at those sites. We discuss these findings in relation to several environmental gradients within the cave (i.e., differences in toxicity and lighting conditions), but we also tentatively argue that the annual fish harvest during a religious ceremony (La Pesca) locally diminishes competition (and possibly, cannibalism by large adults), which is followed by a phase of overcompensation of fish densities. PMID:25083351

  10. ‘Transient’ Genetic Suppression Facilitates Generation of Hexose Transporter Null Mutants in Leishmania mexicana

    PubMed Central

    Feng, Xiuhong; Rodriguez-Contreras, Dayana; Polley, Tamsen; Lye, Lon-Fye; Scott, David; Burchmore, Richard J.S.; Beverley, Stephen M.; Landfear, Scott M.

    2012-01-01

    Summary The genome of Leishmania mexicana encompasses a cluster of three glucose transporter genes designated LmxGT1, LmxGT2, and LmxGT3. Functional and genetic studies of a cluster null mutant (Δlmxgt1-3) have dissected the roles of these proteins in Leishmania metabolism and virulence. However, null mutants were recovered at very low frequency, and comparative genome hybridizations revealed that Δlmxgt1-3 mutants contained a linear extrachromosomal 40 kb amplification of a region on chromosome 29 not amplified in WT parasites. These data suggested a model where this 29–40k amplicon encoded a second site suppressor contributing to parasite survival in the absence of GT1-3 function. To test this, we quantified the frequency of recovery of knockouts in the presence of individual overexpressed ORFs covering the 29–40k amplicon. The data mapped the suppressor activity to PIFTC3, encoding a component of the intraflagellar transport pathway. We discuss possible models by which PIFTC3 might act to facilitate loss of GTs specifically. Surprisingly, by plasmid segregation we showed that continued PIFTC3 overexpression was not required for Δlmxgt1-3 viability. These studies provide the first evidence that genetic suppression can occur by providing critical biological functions transiently. This novel form of genetic suppression may extend to other genes, pathways and organisms. PMID:23170981

  11. Variation in ectomycorrhizal fungal communities associated with Oreomunnea mexicana (Juglandaceae) in a Neotropical montane forest.

    PubMed

    Corrales, Adriana; Arnold, A Elizabeth; Ferrer, Astrid; Turner, Benjamin L; Dalling, James W

    2016-01-01

    Neotropical montane forests are often dominated by ectomycorrhizal (EM) tree species, yet the diversity of their EM fungal communities remains poorly explored. In lower montane forests in western Panama, the EM tree species Oreomunnea mexicana (Juglandaceae) forms locally dense populations in forest otherwise characterized by trees that form arbuscular mycorrhizal (AM) associations. The objective of this study was to compare the composition of EM fungal communities associated with Oreomunnea adults, saplings, and seedlings across sites differing in soil fertility and the amount and seasonality of rainfall. Analysis of fungal nrITS DNA (nuclear ribosomal internal transcribed spacers) revealed 115 EM fungi taxa from 234 EM root tips collected from adults, saplings, and seedlings in four sites. EM fungal communities were equally species-rich and diverse across Oreomunnea developmental stages and sites, regardless of soil conditions or rainfall patterns. However, ordination analysis revealed high compositional turnover between low and high fertility/rainfall sites located ca. 6 km apart. The EM fungal community was dominated by Russula (ca. 36 taxa). Cortinarius, represented by 14 species and previously reported to extract nitrogen from organic sources under low nitrogen availability, was found only in low fertility/high rainfall sites. Phylogenetic diversity analyses of Russula revealed greater evolutionary distance among taxa found on sites with contrasting fertility and rainfall than was expected by chance, suggesting that environmental differences among sites may be important in structuring EM fungal communities. More research is needed to evaluate whether EM fungal taxa associated with Oreomunnea form mycorrhizal networks that might account for local dominance of this tree species in otherwise diverse forest communities. PMID:25940407

  12. A galactosyl(alpha 1-3)mannose epitope on phospholipids of Leishmania mexicana and L. braziliensis is recognized by trypanosomatid-infected human sera.

    PubMed Central

    Avila, J L; Rojas, M

    1990-01-01

    An immunoglobulin M antibody reactive with galactosyl(alpha 1-3)mannose [Gal(alpha 1-3)Man] residues present on phospholipids extracted from Leishmania mexicana and L. braziliensis was found to be present in high titer in the serum of every normal individual studied. Periodate oxidation, acid hydrolysis, or acetylation suppressed immunoreactivity, suggesting that an oligosaccharide chain was responsible for antibody binding. Interaction occurs only with alpha-Gal terminal residues, since treatment of purified glycophospholipids with alpha-galactosidase but not with beta-galactosidase abolished it. Antibody bound to galactosyl(alpha 1-3)galactose-linked synthetic antigens but did not bind to the same residues present in rabbit, rat, and guinea pig erythrocytes or in murine laminin. Antigen-antibody binding was strongly blocked with Gal(alpha 1-3)Man and Gal(beta 1-4)Man. These results plus inhibition studies with several oligosaccharides suggest that they are indeed different from antibodies against the galactosyl(alpha 1-3)galactose residue. Anti-Gal(alpha 1-3)Man antibody values were significantly elevated in 89% of patients with diffuse cutaneous leishmaniasis, 84% of patients with localized cutaneous leishmaniasis, 69% of patients with mucocutaneous leishmaniasis, and 44 and 62% of patients with Trypanosoma cruzi or T. rangeli infection, respectively, but not in patients with 15 other different infectious and inflammatory diseases. Anti-Gal(alpha 1-3)Man antibody readily absorbed to American Leishmania and Trypanosoma culture forms, suggesting a surface membrane localization of reactive epitope. Gal(alpha 1-3)Man-bearing glycophospholipid was easily extracted from American Leishmania promastigotes and T. cruzi trypomastigotes as well as from American Trypanosoma culture forms. The possibility that this antibody arises against parasitic glycophospholipid-linked Gal(alpha 1-3)Man terminal residues is proposed. PMID:1696285

  13. Outbreeding and lack of temporal genetic structure in a drone congregation of the neotropical stingless bee Scaptotrigona mexicana.

    PubMed

    Mueller, Matthias Y; Moritz, Robin Fa; Kraus, F Bernhard

    2012-06-01

    Drone aggregations are a widespread phenomenon in many stingless bee species (Meliponini), but the ultimate and proximate causes for their formation are still not well understood. One adaptive explanation for this phenomenon is the avoidance of inbreeding, which is especially detrimental for stingless bees due to the combined effects of the complementary sex-determining system and the small effective population size caused by eusociality and monandry. We analyzed the temporal genetic dynamics of a drone aggregation of the stingless bee Scaptotrigona mexicana with microsatellite markers over a time window of four weeks. We estimated the drones of the aggregation to originate from a total of 55 colonies using sibship re-construction. There was no detectable temporal genetic differentiation or sub-structuring in the aggregation. Most important, we could exclude all colonies in close proximity of the aggregation as origin of the drones in the aggregation, implicating that they originate from more distant colonies. We conclude that the diverse genetic composition and the distant origin of the drones of the S. mexicana drone congregation provides an effective mechanism to avoid mating among close relatives. PMID:22833802

  14. Morphological and molecular identification of the ectomycorrhizal association of Lactarius fumosibrunneus and Fagus grandifolia var. mexicana trees in eastern Mexico.

    PubMed

    Garay-Serrano, Edith; Bandala, Victor Manuel; Montoya, Leticia

    2012-11-01

    A population of Fagus grandifolia var. mexicana (covering ca. 4.7 ha) is established in a montane cloud forest refuge at Acatlan Volcano in eastern Mexico (Veracruz State), and it represents one of only ten populations of this species known to occur in the country (each stand covers ca. 2-35 ha in extension) and one of the southernmost in the continent. Sporocarps of several ectomycorrhizal macrofungi have been observed in the area, and among them, individuals of the genus Lactarius are common in the forest. However, the morphological and molecular characterization of ectomycorrhizae is still in development. Currently, two species of Lactarius have been previously documented in the area. Through the phylogenetic analysis of the internal transcribed spacer (ITS) region from basidiomes and ectomycorrhizae, we identified the Lactarius fumosibrunneus ectomycorrhiza. The host, F. grandifolia var. mexicana, was determined comparing the amplified ITS sequence from ectomycorrhizal root tips in the GenBank database with Basic Local Alignment Search Tool. The mycorrhizal system of L. fumosibrunneus is monopodial-pyramidal, characterized by its shiny, white to silver and pruinose surface, secreting a white latex when damaged, composed of three plectenchymatous mantle layers, with diverticulated terminal elements at the outer mantle. It lacks emanating hyphae, rhizomorphs, and sclerotia. A detailed morphological and anatomical description, illustrations, and photographs of the ectomycorrhiza are presented. The comparison of L. fumosibrunneus and other Lactarius belonging to subgenus Plinthogalus is presented. PMID:22402818

  15. High precision during food recruitment of experienced (reactivated) foragers in the stingless bee Scaptotrigona mexicana (Apidae, Meliponini)

    NASA Astrophysics Data System (ADS)

    Sánchez, Daniel; Nieh, James C.; Hénaut, Yann; Cruz, Leopoldo; Vandame, Rémy

    Several studies have examined the existence of recruitment communication mechanisms in stingless bees. However, the spatial accuracy of location-specific recruitment has not been examined. Moreover, the location-specific recruitment of reactivated foragers, i.e., foragers that have previously experienced the same food source at a different location and time, has not been explicitly examined. However, such foragers may also play a significant role in colony foraging, particularly in small colonies. Here we report that reactivated Scaptotrigona mexicana foragers can recruit with high precision to a specific food location. The recruitment precision of reactivated foragers was evaluated by placing control feeders to the left and the right of the training feeder (direction-precision tests) and between the nest and the training feeder and beyond it (distance-precision tests). Reactivated foragers arrived at the correct location with high precision: 98.44% arrived at the training feeder in the direction trials (five-feeder fan-shaped array, accuracy of at least +/-6° of azimuth at 50 m from the nest), and 88.62% arrived at the training feeder in the distance trials (five-feeder linear array, accuracy of at least +/-5 m or +/-10% at 50 m from the nest). Thus, S. mexicana reactivated foragers can find the indicated food source at a specific distance and direction with high precision, higher than that shown by honeybees, Apis mellifera, which do not communicate food location at such close distances to the nest.

  16. Outbreeding and lack of temporal genetic structure in a drone congregation of the neotropical stingless bee Scaptotrigona mexicana

    PubMed Central

    Mueller, Matthias Y; Moritz, Robin FA; Kraus, F Bernhard

    2012-01-01

    Drone aggregations are a widespread phenomenon in many stingless bee species (Meliponini), but the ultimate and proximate causes for their formation are still not well understood. One adaptive explanation for this phenomenon is the avoidance of inbreeding, which is especially detrimental for stingless bees due to the combined effects of the complementary sex-determining system and the small effective population size caused by eusociality and monandry. We analyzed the temporal genetic dynamics of a drone aggregation of the stingless bee Scaptotrigona mexicana with microsatellite markers over a time window of four weeks. We estimated the drones of the aggregation to originate from a total of 55 colonies using sibship re-construction. There was no detectable temporal genetic differentiation or sub-structuring in the aggregation. Most important, we could exclude all colonies in close proximity of the aggregation as origin of the drones in the aggregation, implicating that they originate from more distant colonies. We conclude that the diverse genetic composition and the distant origin of the drones of the S. mexicana drone congregation provides an effective mechanism to avoid mating among close relatives. PMID:22833802

  17. PPAR activation induces M1 macrophage polarization via cPLA₂-COX-2 inhibition, activating ROS production against Leishmania mexicana.

    PubMed

    Díaz-Gandarilla, J A; Osorio-Trujillo, C; Hernández-Ramírez, V I; Talamás-Rohana, P

    2013-01-01

    Defence against Leishmania depends upon Th1 inflammatory response and, a major problem in susceptible models, is the turnoff of the leishmanicidal activity of macrophages with IL-10, IL-4, and COX-2 upregulation, as well as immunosuppressive PGE2, all together inhibiting the respiratory burst. Peroxisome proliferator-activated receptors (PPAR) activation is responsible for macrophages polarization on Leishmania susceptible models where microbicide functions are deactivated. In this paper, we demonstrated that, at least for L. mexicana, PPAR activation, mainly PPAR γ , induced macrophage activation through their polarization towards M1 profile with the increase of microbicide activity against intracellular pathogen L. mexicana. PPAR activation induced IL-10 downregulation, whereas the production of proinflammatory cytokines such as TNF- α , IL-1 β , and IL-6 remained high. Moreover, PPAR agonists treatment induced the deactivation of cPLA2-COX-2-prostaglandins pathway together with an increase in TLR4 expression, all of whose criteria meet the M1 macrophage profile. Finally, parasite burden, in treated macrophages, was lower than that in infected nontreated macrophages, most probably associated with the increase of respiratory burst in these treated cells. Based on the above data, we conclude that PPAR agonists used in this work induces M1 macrophages polarization via inhibition of cPLA2 and the increase of aggressive microbicidal activity via reactive oxygen species (ROS) production. PMID:23555077

  18. Toxic hydrogen sulphide and dark caves: pronounced male life-history divergence among locally adapted Poecilia mexicana (Poeciliidae).

    PubMed

    Riesch, R; Plath, M; Schlupp, I

    2011-03-01

    Chronic environmental stress is known to induce evolutionary change. Here, we assessed male life-history trait divergence in the neotropical fish Poecilia mexicana from a system that has been described to undergo incipient ecological speciation in adjacent, but reproductively isolated toxic/nontoxic and surface/cave habitats. Examining both field-caught and common garden-reared specimens, we investigated the extent of differentiation and plasticity of life-history strategies employed by male P. mexicana. We found strong site-specific life-history divergence in traits such as fat content, standard length and gonadosomatic index. The majority of site-specific life-history differences were also expressed under common garden-rearing conditions. We propose that apparent conservatism of male life histories is the result of other (genetically based) changes in physiology and behaviour between populations. Together with the results from previous studies, this is strong evidence for local adaptation as a result of ecologically based divergent selection. PMID:21159007

  19. Two Lactarius species associated with a relict Fagus grandifolia var. mexicana population in a Mexican montane cloud forest.

    PubMed

    Montoya, L; Haug, I; Bandala, V M

    2010-01-01

    Ectomycorrhizal (EM) fleshy fungi are being monitored in a population of Fagus grandifolia var. mexicana persisting in a montane cloud forest refuge on a volcano in a subtropical region of central Veracruz (eastern Mexico). The population of Fagus studied represents one of the 10 recognized forest fragments still housing this tree genus in Mexico. This is the first attempt to document EM fungi associated with this tree species in Mexico. We present evidence of the ectomycorrhizal symbiosis for Lactarius badiopallescens and L. cinereus with this endemic tree. Species identification of Lactarius on Fagus grandifolia var. mexicana was based on the comparison of DNAsequences (ITS rDNA) of spatiotemporally co-occurring basidiomes and EM root tips. The host of the EM tips was identified by comparison of the large subunit of the ribulose-bisphosphate carboxylase gene (rbcL). The occurrence of Lactarius badiopallescens and L. cinereus populations in the area of study represent the southernmost record known to date of these two species in North America and are new for the Neotropical Lactarius mycota. Descriptions coupled with illustrations of macro- and micromorphological features of basidiomes as well as photographs of ectomycorrhizas are presented. PMID:20120238

  20. The role of androgens in species recognition and sperm production in Atlantic mollies (Poecilia mexicana).

    PubMed

    Gabor, Caitlin R; Aspbury, Andrea S; Ma, Jacqueline; Nice, Chris C

    2012-02-01

    Much is known about the role of hormones in the regulation of vertebrate mating behavior, including receptivity, and several components of mate choice. Hormones may modulate reproductive behavior in such a way to increase or decrease the individual's motivation, and therefore hormones may be important in mediating behavior associated with reproductive isolation. The mating complex of the all female gynogenetic Amazon mollies, Poecilia formosa, and their parental species (sailfin mollies, P. latipinna, and Atlantic mollies, P. mexicana) is a model system for studying ultimate mechanisms of species recognition. However, proximate mechanisms, such as variation in hormone levels, and the effect of hormones on sperm production have not been extensively examined. We predict that one or more of the sex steroid hormones in teleost fish (11-ketotestosterone (KT), testosterone (T), and estradiol (E)) will play a role in species recognition (during mate choice and/or sperm priming) for Atlantic mollies (the maternal parental species) that are sympatric with Amazon mollies. We sequentially paired male Atlantic mollies with female conspecifics and Amazon mollies and obtained water-borne hormone samples before and after mating for all fish. We measured circulating KT, T, and E from the water samples. Although we did not find an overall KT response to mating with conspecifics as has been found previously in sailfin mollies, male Atlantic mollies that mated more with conspecific females had lower postmating T levels. Additionally, males attempted to mate more with conspecific females that had lower postmating E levels, but attempted to mate more with Amazon mollies that had higher postmating KT levels. We also examined the effect of KT on sperm priming (a mechanism of premating mate choice), and found that KT levels of male Atlantic mollies prior to mating are correlated with the sperm priming response when males were paired with conspecific females, but this correlation was not

  1. DIABETES MELLITUS COMO FACTOR DE RIESGO DE DEMENCIA EN LA POBLACIÓN ADULTA MAYOR MEXICANA

    PubMed Central

    Silvia, Mejía-Arango; Clemente, y Zúñiga-Gil

    2012-01-01

    Introduccion La diabetes mellitus y las demencias constituyen dos problemas crecientes de salud entre la población adulta mayor del mundo y en particular de los paises en desarrollo. Hacen falta estudios longitudinales sobre el papel de la diabetes como factor de riesgo para demencia. Objetivo Determinar el riesgo de demencia en sujetos Mexicanos con diabetes mellitus tipo 2. Materiales y Metodos Los sujetos diabéticos libres de demencia pertenecientes al Estudio Nacional de Salud y Envejecimiento en México fueron evaluados a los dos años de la línea de base. Se estudió el papel de los factores sociodemográficos, de otras comorbilidades y del tipo de tratamiento en la conversión a demencia. Resultados Durante la línea de base 749 sujetos (13.8%) tuvieron diabetes. El riesgo de desarrollar demencia en estos individuos fue el doble (RR, 2.08 IC 95%, 1.59–2.73). Se encontró un riesgo mayor en individuos de 80 años y más (RR 2.44 IC 95%, 1.46–4.08), en los hombres (RR, 2.25 IC 95%, 1.46–3.49) y en sujetos con nivel educativo menor de 7 años. El estar bajo tratamiento con insulina incrementó el riesgo de demencia (RR, 2.83, IC 95%, 1.58–5.06). Las otras comorbilidades que aumentaron el riesgo de demencia en los pacientes diabéticos fueron la hipertensión (RR, 2.75, IC 95%, 1.86–4.06) y la depresión (RR, 3.78, 95% IC 2.37–6.04). Conclusión Los sujetos con diabetes mellitus tienen un riesgo mayor de desarrollar demencia, La baja escolaridad y otras comorbilidades altamente prevalentes en la población Mexicana contribuyen a la asociación diabetes-demencia. PMID:21948010

  2. Scanning and three-dimensional electron microscopy methods for the study of Trypanosoma brucei and Leishmania mexicana flagella

    PubMed Central

    Gluenz, Eva; Wheeler, Richard John; Hughes, Louise; Vaughan, Sue

    2015-01-01

    Three-dimensional electron microscopy tools have revolutionized our understanding of cell structure and molecular complexes in biology. Here, we describe methods for studying flagellar ultrastructure and biogenesis in two unicellular parasites—Trypanosoma brucei and Leishmania mexicana. We describe methods for the preparation of these parasites for scanning electron microscopy cellular electron tomography, and serial block face scanning electron microscopy (SBFSEM). These parasites have a highly ordered cell shape and form, with a defined positioning of internal cytoskeletal structures and organelles. We show how knowledge of these can be used to dissect cell cycles in both parasites and identify the old flagellum from the new in T. brucei. Finally, we demonstrate the use of SBFSEM three-dimensional models for analysis of individual whole cells, demonstrating the excellent potential this technique has for future studies of mutant cell lines. PMID:25837406

  3. Leishmania species: mechanisms of complement activation by five strains of promastigotes.

    PubMed

    Mosser, D M; Burke, S K; Coutavas, E E; Wedgwood, J F; Edelson, P J

    1986-12-01

    The interaction of fresh serum with promastigotes of Leishmania major, L. donovani, L. mexicana mexicana, L. mexicana amazonensis, and L. braziliensis guyanensis results in lysis of all strains tested with either fresh human or guinea pig serum at 37 C for 30 min. Lysis does not occur in the cold and requires divalent cations and complement that is active hemolytically. Serum deficient in the eighth component of complement is not lytic. Lysis of L. major, L. mexicana, and L. braziliensis proceeds fully in human serum containing EGTA/Mg2+ or in guinea pig serum deficient in the fourth complement component. These species consume only small amounts of C4 from human serum and do not require calcium to optimally bind C3. The data indicate that all are activators of the alternative complement pathway and that the classical pathway is not required for the lysis of these organisms. Promastigotes of L. donovani, in contrast, activate the classical pathway. The presence of calcium is required for both optimal C3 binding and parasite lysis, and L. donovani promastigotes consume C4 when incubated in human serum. In high concentrations, human serum agglutinates all tested Leishmania spp. The agglutinating factor does not require divalent cations, is heat stable, and works at 4 C, suggesting that it is an antibody. This "naturally occurring" antibody cross reacts with all Leishmania spp. and agglutinates them. The adsorption of serum with any Leishmania species or with beads that are Protein A coated, removes the agglutinogen. This factor causes a slight enhancement in alternative pathway activation by L. major and mediates the classical activation by L. donovani. In adsorbed serum, L. donovani promastigotes only weakly activate the alternative complement pathway. Increased concentrations of adsorbed serum are therefore necessary for lysis to proceed. The titer can be partially restored by the addition of heat inactivated serum. Using purified components of the classical cascade

  4. The role of Revista Mexicana de Astronomía y Astrofísica Serie de Conferencias in the world of astronomy

    NASA Astrophysics Data System (ADS)

    Torres-Peimbert, Silvia; Allen, Christine

    2015-08-01

    Forty years ago Revista Mexicana de Astronomía y Astrofísica decided to include the proceedings of astronomical meetings in Mexico and Latin America. In 1995 it became necessary to found the Serie de Conferencias to better differentiate proceedings from refereed papers.So far there have been 58 astronomical meetings published and there are several more in store for the coming years

  5. Synthetic fragments of antigenic lipophosphoglycans from Leishmania major and Leishmania mexicana and their use for characterisation of the Leishmania elongating alpha-D-mannopyranosylphosphate transferase.

    PubMed

    Higson, Adrian P; Ross, Andrew J; Tsvetkov, Yury E; Routier, Françoise H; Sizova, Olga V; Ferguson, Michael A J; Nikolaev, Andrei V

    2005-03-18

    The phosphorylated branched heptasaccharides 7 and 8, the octasaccharide 9 and the phosphorylated trisaccharides 5 and 6, which are fragments of the phosphoglycan portion of the surface lipophosphoglycans from Leishmania mexicana (5) or L. major (6-9), were synthesised by using the glycosyl hydrogenphosphonate method for the preparation of phosphodiester bridges. The compounds were tested as acceptor substrates/putative inhibitors for the Leishmania elongating alpha-D-mannosylphosphate transferase. PMID:15685582

  6. Comparison of small mammal prevalence of Leishmania (Leishmania) mexicana in five foci of cutaneous leishmaniasis in the State of Campeche, Mexico.

    PubMed

    Van Wynsberghe, N R; Canto-Lara, S B; Sosa-Bibiano, E I; Rivero-Cárdenas, N A; Andrade-Narváez, F J

    2009-01-01

    In the Yucatan Peninsula of Mexico, 95% of the human cases of Cutaneous Leishmaniasis are caused by Leishmania (Leishmania) mexicana with an incidence rate of 5.08 per 100,000 inhabitants. Transmission is limited to the winter months (November to March). One study on wild rodents has incriminated Ototylomys phyllotis and Peromyscus yucatanicus as primary reservoirs of L. (L.) mexicana in the focus of La Libertad, Campeche. In the present study, the prevalence of both infection and disease caused by L. (L.) mexicana in small terrestrial mammals were documented during five transmission seasons (1994-2004) in five foci of Leishmaniasis in the state of Campeche. Foci separated by only 100 km, with similar relative abundances of small mammals, were found to differ significantly in their prevalence of both symptoms and infection. Transmission rates and reservoir species seemed to change in space as well as in time which limited the implementation of effective control measures of the disease even in a small endemic area such as the south of the Yucatan Peninsula. PMID:19390737

  7. Sex-specific local life-history adaptation in surface- and cave-dwelling Atlantic mollies (Poecilia mexicana).

    PubMed

    Riesch, Rüdiger; Reznick, David N; Plath, Martin; Schlupp, Ingo

    2016-01-01

    Cavefishes have long been used as model organisms showcasing adaptive diversification, but does adaptation to caves also facilitate the evolution of reproductive isolation from surface ancestors? We raised offspring of wild-caught surface- and cave-dwelling ecotypes of the neotropical fish Poecilia mexicana to sexual maturity in a 12-month common garden experiment. Fish were raised under one of two food regimes (high vs. low), and this was crossed with differences in lighting conditions (permanent darkness vs. 12:12 h light:dark cycle) in a 2 × 2 factorial design, allowing us to elucidate potential patterns of local adaptation in life histories. Our results reveal a pattern of sex-specific local life-history adaptation: Surface molly females had the highest fitness in the treatment best resembling their habitat of origin (high food and a light:dark cycle), and suffered from almost complete reproductive failure in darkness, while cave molly females were not similarly affected in any treatment. Males of both ecotypes, on the other hand, showed only weak evidence for local adaptation. Nonetheless, local life-history adaptation in females likely contributes to ecological diversification in this system and other cave animals, further supporting the role of local adaptation due to strong divergent selection as a major force in ecological speciation. PMID:26960566

  8. Sex-specific local life-history adaptation in surface- and cave-dwelling Atlantic mollies (Poecilia mexicana)

    PubMed Central

    Riesch, Rüdiger; Reznick, David N.; Plath, Martin; Schlupp, Ingo

    2016-01-01

    Cavefishes have long been used as model organisms showcasing adaptive diversification, but does adaptation to caves also facilitate the evolution of reproductive isolation from surface ancestors? We raised offspring of wild-caught surface- and cave-dwelling ecotypes of the neotropical fish Poecilia mexicana to sexual maturity in a 12-month common garden experiment. Fish were raised under one of two food regimes (high vs. low), and this was crossed with differences in lighting conditions (permanent darkness vs. 12:12 h light:dark cycle) in a 2 × 2 factorial design, allowing us to elucidate potential patterns of local adaptation in life histories. Our results reveal a pattern of sex-specific local life-history adaptation: Surface molly females had the highest fitness in the treatment best resembling their habitat of origin (high food and a light:dark cycle), and suffered from almost complete reproductive failure in darkness, while cave molly females were not similarly affected in any treatment. Males of both ecotypes, on the other hand, showed only weak evidence for local adaptation. Nonetheless, local life-history adaptation in females likely contributes to ecological diversification in this system and other cave animals, further supporting the role of local adaptation due to strong divergent selection as a major force in ecological speciation. PMID:26960566

  9. Assessing the importance of four sandfly species (Diptera: Psychodidae) as vectors of Leishmania mexicana in Campeche, Mexico.

    PubMed

    Pech-May, A; Peraza-Herrera, G; Moo-Llanes, D A; Escobedo-Ortegón, J; Berzunza-Cruz, M; Becker-Fauser, I; Montes DE Oca-Aguilar, A C; Rebollar-Téllez, E A

    2016-09-01

    Localized cutaneous leishmaniasis represents a public health problem in many areas of Mexico, especially in the Yucatan Peninsula. An understanding of vector ecology and bionomics is of great importance in evaluations of the transmission dynamics of Leishmania parasites. A field study was conducted in the county of Calakmul, state of Campeche, during the period from November 2006 to March 2007. Phlebotomine sandfly vectors were sampled using Centers for Disease Control light traps, baited Disney traps and Shannon traps. A total of 3374 specimens were captured in the two villages of Once de Mayo (93.8%) and Arroyo Negro (6.1%). In Once de Mayo, the most abundant species were Psathyromyia shannoni, Lutzomyia cruciata, Bichromomyia olmeca olmeca and Psychodopygus panamensis (all: Diptera: Psychodidae). The Shannon trap was by far the most efficient method of collection. The infection rate, as determined by Leishmania mexicana-specific polymerase chain reaction, was 0.3% in Once de Mayo and infected sandflies included Psy. panamensis, B. o. olmeca and Psa. shannoni. There were significant differences in human biting rates across sandfly species and month of sampling. Ecological niche modelling analyses showed an overall overlap of 39.1% for the four species in the whole state of Campeche. In addition, the finding of nine vector-reservoir pairs indicates a potential interaction. The roles of the various sandfly vectors in Calakmul are discussed. PMID:27040367

  10. Choosy males from the underground: male mating preferences in surface- and cave-dwelling Atlantic mollies ( Poecilia mexicana)

    NASA Astrophysics Data System (ADS)

    Plath, Martin; Seggel, Uta; Burmeister, Heike; Heubel, Katja U.; Schlupp, Ingo

    2006-03-01

    Atlantic mollies ( Poecilia mexicana) inhabit a variety of surface habitats, but they also occur in a sulfur cave in southern Mexico. We examined male mate choice relative to female body size in the cave population and in the most closely related surface-dwelling population from a nearby river. Males from both populations were either light- or dark-reared and could choose between two differently sized females either on the basis of visual cues in light or on the basis of solely nonvisual cues in darkness. Sexual preferences were estimated from the degree of association. Cave molly males always showed a preference for the larger female, both in light and in darkness. Among the surface males, only light-reared males showed a preference in the visual cues test, but not in darkness. In a control experiment, we demonstrated that male association preferences directly translate into actual mating preferences. Apparently, using visual cues for mate choice is the ancestral state in this system, and using nonvisual cues has evolved as a novel trait in the cave population. We discuss the evolution of nonvisual male mate choice in the context of changed environmental conditions, namely the absence of light, hypoxia, and toxic hydrogen sulfide in the cave.

  11. Toxic hydrogen sulfide and dark caves: life-history adaptations in a livebearing fish (Poecilia mexicana, Poeciliidae).

    PubMed

    Riesch, Rüdiger; Plath, Martin; Schlupp, Ingo

    2010-05-01

    Life-history traits are very sensitive to extreme environmental conditions, because resources that need to be invested in somatic maintenance cannot be invested in reproduction. Here we examined female life-history traits in the Mexican livebearing fish Poecilia mexicana from a variety of benign surface habitats, a creek with naturally occurring toxic hydrogen sulfide (H2S), a sulfidic cave, and a non-sulfidic cave. Previous studies revealed pronounced genetic and morphological divergence over very small geographic scales in this system despite the absence of physical barriers, suggesting that local adaptation to different combinations of two selection factors, toxicity (H2S) and darkness, is accompanied by very low rates of gene flow. Hence, we investigated life-history divergence between these populations in response to the selective pressures of darkness and/or toxicity. Our main results show that toxicity and darkness both select for (or impose constraints on) the same female trait dynamics: reduced fecundity and increased offspring size. Since reduced fecundity in the sulfur cave population was previously shown to be heritable, we discuss how divergent life-history evolution may promote further ecological divergence: for example, reduced fecundity and increased offspring autonomy are clearly beneficial in extreme environments, but fish with these traits are outcompeted in benign habitats. PMID:20503881

  12. Choosy males from the underground: male mating preferences in surface- and cave-dwelling Atlantic mollies (Poecilia mexicana).

    PubMed

    Plath, Martin; Seggel, Uta; Burmeister, Heike; Heubel, Katja U; Schlupp, Ingo

    2006-03-01

    Atlantic mollies (Poecilia mexicana) inhabit a variety of surface habitats, but they also occur in a sulfur cave in southern Mexico. We examined male mate choice relative to female body size in the cave population and in the most closely related surface-dwelling population from a nearby river. Males from both populations were either light- or dark-reared and could choose between two differently sized females either on the basis of visual cues in light or on the basis of solely nonvisual cues in darkness. Sexual preferences were estimated from the degree of association. Cave molly males always showed a preference for the larger female, both in light and in darkness. Among the surface males, only light-reared males showed a preference in the visual cues test, but not in darkness. In a control experiment, we demonstrated that male association preferences directly translate into actual mating preferences. Apparently, using visual cues for mate choice is the ancestral state in this system, and using nonvisual cues has evolved as a novel trait in the cave population. We discuss the evolution of nonvisual male mate choice in the context of changed environmental conditions, namely the absence of light, hypoxia, and toxic hydrogen sulfide in the cave. PMID:16404589

  13. Toxic hydrogen sulfide and dark caves: phenotypic and genetic divergence across two abiotic environmental gradients in Poecilia mexicana.

    PubMed

    Tobler, Michael; Dewitt, Thomas J; Schlupp, Ingo; García de León, Francisco J; Herrmann, Roger; Feulner, Philine G D; Tiedemann, Ralph; Plath, Martin

    2008-10-01

    Divergent natural selection drives evolutionary diversification. It creates phenotypic diversity by favoring developmental plasticity within populations or genetic differentiation and local adaptation among populations. We investigated phenotypic and genetic divergence in the livebearing fish Poecilia mexicana along two abiotic environmental gradients. These fish typically inhabit nonsulfidic surface rivers, but also colonized sulfidic and cave habitats. We assessed phenotypic variation among a factorial combination of habitat types using geometric and traditional morphometrics, and genetic divergence using quantitative and molecular genetic analyses. Fish in caves (sulfidic or not) exhibited reduced eyes and slender bodies. Fish from sulfidic habitats (surface or cave) exhibited larger heads and longer gill filaments. Common-garden rearing suggested that these morphological differences are partly heritable. Population genetic analyses using microsatellites as well as cytochrome b gene sequences indicate high population differentiation over small spatial scale and very low rates of gene flow, especially among different habitat types. This suggests that divergent environmental conditions constitute barriers to gene flow. Strong molecular divergence over short distances as well as phenotypic and quantitative genetic divergence across habitats in directions classic to fish ecomorphology suggest that divergent selection is structuring phenotypic variation in this system. PMID:18637957

  14. Ser/Thr-rich repetitive motifs as targets for phosphoglycan modifications in Leishmania mexicana secreted acid phosphatase.

    PubMed

    Wiese, M; Ilg, T; Lottspeich, F; Overath, P

    1995-03-15

    The insect stage of the protozoan parasite Leishmania mexicana secretes a phosphomonoesterase in the form of a filamentous complex. The polypeptide subunits of this polymer are modified by phosphoglycans and/or oligomannosyl residues linked to phosphoserine. Based on peptide sequence data of a predominant 100 kDa protein of the filamentous complex, two tandemly arranged, single copy genes, lmsap1 and lmsap2, were cloned and sequenced. lmsap1 predicts a protein with features characteristic of acid phosphatases and a remarkable serine- and threonine-rich region of 32 amino acids close to the C-terminus. In the otherwise identical lmsap2 product, this region is extended to 383 amino acids and is composed of short Ser/Thr-rich repeats. Deletion analysis demonstrates that lmsap1 encodes the major 100 kDa protein of the complex while a minor 200 kDa component is derived from the lmsap2 gene. Null mutants of either gene retain the ability to secrete acid phosphatase filaments, while a deletion of both genes results in Leishmania defective in enzyme formation. The Ser/Thr-rich domains are the targets for phosphoglycan modifications as shown by the expression of secreted fusion proteins composed of these C-terminal regions and the N-terminal domain of a lysosomal acid phosphatase. PMID:7720697

  15. An Evaluation of the Effects of Soil Characteristics on Mitigation and Restoration Involving Blue Elderberry, Sambucus mexicana

    NASA Astrophysics Data System (ADS)

    Koch-Munz, Molly; Holyoak, Marcel

    2008-07-01

    We conducted field surveys and laboratory analyses to test the effects of soil characteristics in habitat mitigation sites and natural sites on the growth and condition of blue elderberry ( Sambucus mexicana), which is the sole host plant for the federally threatened Valley elderberry longhorn beetle ( Desmocerus californicus dimorphus). Thirty mitigation and 16 natural sites were selected throughout the range of the beetle. We found that although plant relative growth rates were comparable between mitigation sites and a natural site, mitigation sites contained substantially less soil nutrients than mitigation sites. Within mitigation sites, elderberry health and growth were positively correlated with the amount of total nitrogen in soils and less strongly with other soil nutrients and soil moisture. Analyses demonstrated reductions in the relative growth rate of elderberry as mitigation sites aged, and that soil nutrients and soil moisture became depleted over time. For mitigation sites, it took approximately seven years to develop basal stem diameters that have been linked to successful beetle colonization. Mitigation sites have smaller shrubs than natural sites and growth slows as mitigation sites age, thus delaying convergence of conditions between natural and mitigation sites. Analyses of soil particle size and whether sites were within the 100-year floodplain (as an indicator of riparian conditions) were inconclusive. We recommend investigating fertilizing and optimum planting densities for elderberry at restoration and mitigation sites, as well as increasing the duration of irrigation and monitoring.

  16. Crystal Structure of Arginase from Leishmania mexicana and Implications for the Inhibition of Polyamine Biosynthesis in Parasitic Infections†

    PubMed Central

    D’Antonio, Edward L.; Ullman, Buddy; Roberts, Sigrid C.; Dixit, Upasna Gaur; Wilson, Mary E.; Hai, Yang; Christianson, David W.

    2013-01-01

    Arginase from parasitic protozoa belonging to the genus Leishmania is a potential drug target for the treatment of leishmaniasis because this binuclear manganese metalloenzyme catalyzes the first committed step in the biosynthesis of polyamines that enable cell growth and survival. The high resolution X-ray crystal structures of the unliganded form of Leishmania mexicana arginase (LmARG) and four inhibitor complexes are now reported. These complexes include the reactive substrate analogue 2(S)-amino-6-boronohexanoic acid (ABH) and the hydroxylated substrate analogue nor-Nω-hydroxy-L-arginine (nor-NOHA), which are the most potent arginase inhibitors known to date. Comparisons of the LmARG structure with that of the archetypal arginase, human arginase I, reveal that all residues important for substrate binding and catalysis are strictly conserved. However, three regions of tertiary structure differ between the parasitic enzyme and the human enzyme corresponding to the G62 – S71, L161 – C172, and I219 – V230 segments of LmARG. Additionally, variations are observed in salt link interactions that stabilize trimer assembly in LmARG. We also report biological studies in which we demonstrate that localization of LmARG to the glycosome, a unique subcellular organelle peculiar to Leishmania and related parasites, is essential for robust pathogenesis. PMID:23583962

  17. Otolith morphology and hearing abilities in cave- and surface-dwelling ecotypes of the Atlantic molly, Poecilia mexicana (Teleostei: Poeciliidae)

    PubMed Central

    Schulz-Mirbach, Tanja; Ladich, Friedrich; Riesch, Rüdiger; Plath, Martin

    2010-01-01

    Cave fish have rarely been investigated with regard to their inner ear morphology, hearing abilities, and acoustic communication. Based on a previous study that revealed morphological differences in the saccular otolith between a cave and two surface populations of Poecilia mexicana, we checked for additional differences in utricular and lagenar otoliths and tested whether different populations have similar hearing sensitivities. We found pronounced differences in the shape of all three otoliths. Otoliths of the saccule and lagena from cave fish differed from those of surface fish in the features of the face oriented towards the sensory epithelium. In addition, otoliths of the utricle and lagena were significantly heavier in cave fish. Auditory sensitivities were measured between 100 and 1500 Hz, utilizing the auditory evoked potential recording technique. We found similar hearing abilities in cave and surface fish, with greatest sensitivity between 200 and 300 Hz. An acoustic survey revealed that neither ecotype produced species-specific sounds. Our data indicate that cave dwelling altered the otolith morphology in Atlantic mollies, probably due to metabolic differences. Different otolith morphology, however, did not affect general auditory sensitivity or acoustic behavior. PMID:20430090

  18. Proteome profiling of the growth phases of Leishmania pifanoi promastigotes in axenic culture reveals differential abundance of immunostimulatory proteins.

    PubMed

    Alcolea, Pedro J; Alonso, Ana; García-Tabares, Francisco; Mena, María del Carmen; Ciordia, Sergio; Larraga, Vicente

    2016-06-01

    Leishmaniasis is a term that encompasses a compendium of neglected tropical diseases caused by dimorphic and digenetic protozoan parasites from the genus Leishmania (Kinetoplastida: Trypanosomatidae). The clinical manifestations of neotropical cutaneous leishmaniasis (NCL) caused by Leishmania pifanoi and other species of the "Leishmania mexicana complex" mainly correspond to anergic diffuse cutaneous leishmaniasis (ADCL), which is the origin of considerable morbidity. Despite the outstanding advances in the characterization of the trypanosomatid genomes and proteomes, the biology of this species has been scarcely explored. However, the close relation of L. pifanoi to the sequenced species L. mexicana and others included in the "L. mexicana complex" allowed us to perform a two-dimension electrophoresis (2DE) approach to the promastigote proteome at the differential expression level. Protein identifications were performed by matrix-assisted laser desorption-ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF). This insight has revealed similarities and differences between L. pifanoi and other species responsible for cutaneous and visceral leishmaniasis. Interestingly, certain proteins that were previously described as immunostimulatory (elongation factor 1β, trypanothione peroxidase, heat shock protein 70, enolase, GDP-forming succinyl-CoA and aldehyde dehydrogenase) are more abundant in the final growth stages of promastigotes (late-logarithmic and/or stationary phase) in the case of L. pifanoi. PMID:26992294

  19. Isolation of Leishmania (L.) mexicana from wild rodents and their possible role in the transmission of localized cutaneous leishmaniasis in the state of Campeche, Mexico.

    PubMed

    Chable-Santos, J B; Van Wynsberghe, N R; Canto-Lara, S B; Andrade-Narvaez, F J

    1995-08-01

    Leishmania parasites were isolated from four of seven small rodent species captured in the State of Campeche, Mexico. Signs and/or symptoms of leishmanial infection were restricted to the tail of the mice. Peromyscus yucatanicus endemic to the peninsula of Yucatan and Oryzomys melanotis were added to the list of carriers of Leishmania. Leishmania (L.) mexicana, characterized by an indirect immunofluorescence antibody method using monoclonal antibodies, was identified in one Sigmodon hispidus and one Or. melanotis. The possible role of the different species as primary reservoirs of Leishmania in the peninsula of Yucatan is discussed. PMID:7677214

  20. Crystal structures of Leishmania mexicana phosphoglycerate mutase suggest a one-metal mechanism and a new enzyme subclass.

    PubMed

    Nowicki, Matthew W; Kuaprasert, Buabarn; McNae, Iain W; Morgan, Hugh P; Harding, Marjorie M; Michels, Paul A M; Fothergill-Gilmore, Linda A; Walkinshaw, Malcolm D

    2009-12-01

    The structures of Leishmania mexicana cofactor-independent phosphoglycerate mutase (Lm iPGAM) crystallised with the substrate 3-phosphoglycerate at high and low cobalt concentrations have been solved at 2.00- and 1.90-A resolutions. Both structures are very similar and the active site contains both 3-phosphoglycerate and 2-phosphoglycerate at equal occupancies (50%). Lm iPGAM co-crystallised with the product 2-phosphoglycerate yields the same structure. Two Co(2+) are coordinated within the active site with different geometries and affinities. The cobalt at the M1 site has a distorted octahedral geometry and is present at 100% occupancy. The M2-site Co(2+) binds with distorted tetrahedral geometry, with only partial occupancy, and coordinates with Ser75, the residue involved in phosphotransfer. When the M2 site is occupied, the side chain of Ser75 adopts a position that is unfavourable for catalysis, indicating that this site may not be occupied under physiological conditions and that catalysis may occur via a one-metal mechanism. The geometry of the M2 site suggests that it is possible for Ser75 to be activated for phosphotransfer by H-bonding to nearby residues rather than by metal coordination. The 16 active-site residues of Lm iPGAM are conserved in the Mn-dependent iPGAM from Bacillus stearothermophilus (33% overall sequence identity). However, Lm iPGAM has an inserted tyrosine (Tyr210) that causes the M2 site to diminish in size, consistent with its reduced metal affinity. Tyr210 is present in trypanosomatid and plant iPGAMs, but not in the enzymes from other organisms, indicating that there are two subclasses of iPGAMs. PMID:19781556

  1. Anticonvulsant and Antioxidant Effects of Tilia americana var. mexicana and Flavonoids Constituents in the Pentylenetetrazole-Induced Seizures

    PubMed Central

    Cárdenas-Rodríguez, Noemí; González-Trujano, María Eva; Aguirre-Hernández, Eva; Ruíz-García, Matilde; Sampieri, Aristides; Coballase-Urrutia, Elvia; Carmona-Aparicio, Liliana

    2014-01-01

    Tilia genus is commonly used around the world for its central nervous system properties; it is prepared as tea and used as tranquilizing, anticonvulsant, and analgesic. In this study, anticonvulsant activity of the Tilia americana var. mexicana inflorescences and leaves was investigated by evaluating organic and aqueous extracts (100, 300, and 600 mg/kg, i.p.) and some flavonoids in the pentylenetetrazole-induced seizures in mice. Moreover, antioxidant effect of these extracts and flavonoids was examined in an in vitro study by using spectrophotometric technique. Significant activity was observed in the methanol extract from inflorescences. An HPLC analysis of the methanol extract from inflorescences and leaves of Tilia allowed demonstrating the respective presence of some partial responsible flavonoid constituents: quercetin (20.09 ± 1.20 μg/mg and 3.39 ± 0.10 μg/mg), rutin (3.52 ± 0.21 μg/mg and 8.94 ± 0.45 μg/mg), and isoquercitrin (1.74 ± 0.01 μg/mg and 1.24 ± 0.13 μg/mg). In addition, significant but different antioxidant properties were obtained among the flavonoids and the extracts investigated. Our results provide evidence of the anticonvulsant activity of Tilia reinforcing its utility for central nervous system diseases whose mechanism of action might involve partial antioxidant effects due to the presence of flavonoids. PMID:25197430

  2. Anticonvulsant and antioxidant effects of Tilia americana var. mexicana and flavonoids constituents in the pentylenetetrazole-induced seizures.

    PubMed

    Cárdenas-Rodríguez, Noemí; González-Trujano, María Eva; Aguirre-Hernández, Eva; Ruíz-García, Matilde; Sampieri, Aristides; Coballase-Urrutia, Elvia; Carmona-Aparicio, Liliana

    2014-01-01

    Tilia genus is commonly used around the world for its central nervous system properties; it is prepared as tea and used as tranquilizing, anticonvulsant, and analgesic. In this study, anticonvulsant activity of the Tilia americana var. mexicana inflorescences and leaves was investigated by evaluating organic and aqueous extracts (100, 300, and 600 mg/kg, i.p.) and some flavonoids in the pentylenetetrazole-induced seizures in mice. Moreover, antioxidant effect of these extracts and flavonoids was examined in an in vitro study by using spectrophotometric technique. Significant activity was observed in the methanol extract from inflorescences. An HPLC analysis of the methanol extract from inflorescences and leaves of Tilia allowed demonstrating the respective presence of some partial responsible flavonoid constituents: quercetin (20.09 ± 1.20 μg/mg and 3.39 ± 0.10 μg/mg), rutin (3.52 ± 0.21 μg/mg and 8.94 ± 0.45 μg/mg), and isoquercitrin (1.74 ± 0.01 μg/mg and 1.24 ± 0.13 μg/mg). In addition, significant but different antioxidant properties were obtained among the flavonoids and the extracts investigated. Our results provide evidence of the anticonvulsant activity of Tilia reinforcing its utility for central nervous system diseases whose mechanism of action might involve partial antioxidant effects due to the presence of flavonoids. PMID:25197430

  3. Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8

    PubMed Central

    Judice, Wagner A. S.; Manfredi, Marcella A.; Souza, Gerson P.; Sansevero, Thiago M.; Almeida, Paulo C.; Shida, Cláudio S.; Gesteira, Tarsis F.; Juliano, Luiz; Westrop, Gareth D.; Sanderson, Sanya J.; Coombs, Graham H.; Tersariol, Ivarne L. S.

    2013-01-01

    Background Cysteine protease B is considered crucial for the survival and infectivity of the Leishmania in its human host. Several microorganism pathogens bind to the heparin-like glycosaminoglycans chains of proteoglycans at host-cell surface to promote their attachment and internalization. Here, we have investigated the influence of heparin upon Leishmania mexicana cysteine protease rCPB2.8 activity. Methodology/Principal Findings The data analysis revealed that the presence of heparin affects all steps of the enzyme reaction: (i) it decreases 3.5-fold the k1 and 4.0-fold the k−1, (ii) it affects the acyl-enzyme accumulation with pronounced decrease in k2 (2.7-fold), and also decrease in k3 (3.5-fold). The large values of ΔG  =  12 kJ/mol for the association and dissociation steps indicate substantial structural strains linked to the formation/dissociation of the ES complex in the presence of heparin, which underscore a conformational change that prevents the diffusion of substrate in the rCPB2.8 active site. Binding to heparin also significantly decreases the α-helix content of the rCPB2.8 and perturbs the intrinsic fluorescence emission of the enzyme. The data strongly suggest that heparin is altering the ionization of catalytic (Cys25)-S−/(His163)-Im+ H ion pair of the rCPB2.8. Moreover, the interaction of heparin with the N-terminal pro-region of rCPB2.8 significantly decreased its inhibitory activity against the mature enzyme. Conclusions/Significance Taken together, depending on their concentration, heparin-like glycosaminoglycans can either stimulate or antagonize the activity of cysteine protease B enzymes during parasite infection, suggesting that this glycoconjugate can anchor parasite cysteine protease at host cell surface. PMID:24278253

  4. Unique evolutionary trajectories in repeated adaptation to hydrogen sulphide-toxic habitats of a neotropical fish (Poecilia mexicana).

    PubMed

    Pfenninger, Markus; Patel, Simit; Arias-Rodriguez, Lenin; Feldmeyer, Barbara; Riesch, Rüdiger; Plath, Martin

    2015-11-01

    Replicated ecological gradients are prime systems to study processes of molecular evolution underlying ecological divergence. Here, we investigated the repeated adaptation of the neotropical fish Poecilia mexicana to habitats containing toxic hydrogen sulphide (H2 S) and compared two population pairs of sulphide-adapted and ancestral fish by sequencing population pools of >200 individuals (Pool-Seq). We inferred the evolutionary processes shaping divergence and tested the hypothesis of increase of parallelism from SNPs to molecular pathways. Coalescence analyses showed that the divergence occurred in the face of substantial bidirectional gene flow. Population divergence involved many short, widely dispersed regions across the genome. Analyses of allele frequency spectra suggest that differentiation at most loci was driven by divergent selection, followed by a selection-mediated reduction of gene flow. Reconstructing allelic state changes suggested that selection acted mainly upon de novo mutations in the sulphide-adapted populations. Using a corrected Jaccard index to quantify parallel evolution, we found a negligible proportion of statistically significant parallel evolution of Jcorr  = 0.0032 at the level of SNPs, divergent genome regions (Jcorr  = 0.0061) and genes therein (Jcorr  = 0.0091). At the level of metabolic pathways, the overlap was Jcorr  = 0.2545, indicating increasing parallelism with increasing level of biological integration. The majority of pathways contained positively selected genes in both sulphide populations. Hence, adaptation to sulphidic habitats necessitated adjustments throughout the genome. The largely unique evolutionary trajectories may be explained by a high proportion of de novo mutations driving the divergence. Our findings favour Gould's view that evolution is often the unrepeatable result of stochastic events with highly contingent effects. PMID:26405850

  5. Comparative analysis of the gonadal transcriptomes of the all-female species Poecilia formosa and its maternal ancestor Poecilia mexicana

    PubMed Central

    2014-01-01

    Background The Amazon molly, Poecilia formosa (Teleostei: Poeciliinae) is an unisexual, all-female species. It evolved through the hybridisation of two closely related sexual species and exhibits clonal reproduction by sperm dependent parthenogenesis (or gynogenesis) where the sperm of a parental species is only used to activate embryogenesis of the apomictic, diploid eggs but does not contribute genetic material to the offspring. Here we provide and describe the first de novo assembled transcriptome of the Amazon molly in comparison with its maternal ancestor, the Atlantic molly Poecilia mexicana. The transcriptome data were produced through sequencing of single end libraries (100 bp) with the Illumina sequencing technique. Results 83,504,382 reads for the Amazon molly and 81,625,840 for the Atlantic molly were assembled into 127,283 and 78,961 contigs for the Amazon molly and the Atlantic molly, respectively. 63% resp. 57% of the contigs could be annotated with gene ontology terms after sequence similarity comparisons. Furthermore, we were able to identify genes normally involved in reproduction and especially in meiosis also in the transcriptome dataset of the apomictic reproducing Amazon molly. Conclusions We assembled and annotated the transcriptome of a non-model organism, the Amazon molly, without a reference genome (de novo). The obtained dataset is a fundamental resource for future research in functional and expression analysis. Also, the presence of 30 meiosis-specific genes within a species where no meiosis is known to take place is remarkable and raises new questions for future research. PMID:24742317

  6. Chemical compositions and antimicrobial and antioxidant activities of the essential oils from Magnolia grandiflora, Chrysactinia mexicana, and Schinus molle found in northeast Mexico.

    PubMed

    Guerra-Boone, Laura; Alvarez-Román, Rocío; Salazar-Aranda, Ricardo; Torres-Cirio, Anabel; Rivas-Galindo, Verónica Mayela; Waksman de Torres, Noemí; González González, Gloria María; Pérez-López, Luis Alejandro

    2013-01-01

    The essential oils from Magnolia grandiflora and Chrysactinia mexicana leaves, and from Schinus molle leaves and fruit, were characterized by gas chromatography/flame-ionization detection and gas chromatography/mass spectrometry. Twenty-eight compounds from M. grandiflora leaves were identified (representing 93.6% of the total area of the gas chromatogram), with the major component being bornyl acetate (20.9%). Colorless and yellow oils were obtained from the C. mexicana leaves with 18 (86.7%) and 11 (100%) compounds identified, respectively. In both fractions, the principal component was sylvestrene (36.8% and 41.1%, respectively). The essential oils ofS. molle leaves and fruit were each separated into colorless and yellow fractions, in which 14 (98.2) and 20 (99.8%) compounds were identified. The main component was alpha-phellandrene in all fractions (between 32.8% and 45.0%). The M. grandiflora oil displayed antifungal activity against five dermatophyte strains. The oils from S. molle and M. grandiflora leaves had antimicrobial activity against Staphylococcus aureus and Streptococcus pyogenes, which cause skin infections that potentially may lead to sepsis. However, the antioxidant activities of all oils were small (half maximal effective concentration values >250 microg/mL). PMID:23472479

  7. PPAR Activation Induces M1 Macrophage Polarization via cPLA2-COX-2 Inhibition, Activating ROS Production against Leishmania mexicana

    PubMed Central

    Díaz-Gandarilla, J. A.; Osorio-Trujillo, C.; Hernández-Ramírez, V. I.; Talamás-Rohana, P.

    2013-01-01

    Defence against Leishmania depends upon Th1 inflammatory response and, a major problem in susceptible models, is the turnoff of the leishmanicidal activity of macrophages with IL-10, IL-4, and COX-2 upregulation, as well as immunosuppressive PGE2, all together inhibiting the respiratory burst. Peroxisome proliferator-activated receptors (PPAR) activation is responsible for macrophages polarization on Leishmania susceptible models where microbicide functions are deactivated. In this paper, we demonstrated that, at least for L. mexicana, PPAR activation, mainly PPARγ, induced macrophage activation through their polarization towards M1 profile with the increase of microbicide activity against intracellular pathogen L. mexicana. PPAR activation induced IL-10 downregulation, whereas the production of proinflammatory cytokines such as TNF-α, IL-1β, and IL-6 remained high. Moreover, PPAR agonists treatment induced the deactivation of cPLA2-COX-2-prostaglandins pathway together with an increase in TLR4 expression, all of whose criteria meet the M1 macrophage profile. Finally, parasite burden, in treated macrophages, was lower than that in infected nontreated macrophages, most probably associated with the increase of respiratory burst in these treated cells. Based on the above data, we conclude that PPAR agonists used in this work induces M1 macrophages polarization via inhibition of cPLA2 and the increase of aggressive microbicidal activity via reactive oxygen species (ROS) production. PMID:23555077

  8. Ursolic acid from Agastache mexicana aerial parts produces antinociceptive activity involving TRPV1 receptors, cGMP and a serotonergic synergism.

    PubMed

    Verano, Jazmín; González-Trujano, Ma Eva; Déciga-Campos, Myrna; Ventura-Martínez, Rosa; Pellicer, Francisco

    2013-09-01

    Agastache mexicana is a plant that has long been used in large demands in Mexican folk medicine to treat anxiety, insomnia and pain, among others affections. Chromatographic technique was used to identify ursolic acid (UA), 130.7 mg/g and 20.3 mg/g, as an antinociceptive active compound identified in ethyl acetate and methanol extracts of A. mexicana aerial parts, respectively. Temporal course curves of the antinociceptive response demonstrated a dose-dependent and significant activity of UA (1 to 100 mg/kg, i.p.) with an ED50=2 mg/kg in comparison to the efficacy of diclofenac (1 or 30 to 100 mg/kg, i.p.), a non-steroidal anti-inflammatory drug, with an ED50=11.56 mg/kg. The antinociceptive response consisted in the reduction of abdominal constrictions induced with 1% acetic acid in mice. Similarly, UA at 2 mg/kg produced significant antinociception in the intracolonic administration of 0.3% capsaicin (a TRPV1 agonist) in mice. It has been reported the inhibition produced by UA on the calcium-flux induced by capsaicin on TRPV1 receptor suggesting the antagonistic activity of this receptor. Finally, an ED50=44 mg/kg was calculated in the neurogenic and inflammatory nociception induced in the formalin test in rats. The antinociceptive response of UA in the formalin test was not modified in presence of naloxone, flumazenil or L-arginine. Nevertheless, it was reverted in presence of 1-H-(1,2,4)-oxadiazolo(4,2-a)quinoxalin-1-one (ODQ, an inhibitor of soluble guanylyl cyclase) and increased in presence of N(G)-L-nitro-arginine methyl ester (L-NAME, inhibitor of nitric oxide synthase), theophylline (inhibitor of phosphodiesterase) and WAY100635 (an antagonist of 5-HT1A receptors). Current results provide evidence that the antinociceptive response of A. mexicana depends in part on the presence of UA. Moreover, this triterpene may exerts its antinociceptive effect mediated by the presence of cGMP and an additive synergism with 5HT1A receptors, but also an antagonistic

  9. Down-Regulation of TLR and JAK/STAT Pathway Genes Is Associated with Diffuse Cutaneous Leishmaniasis: A Gene Expression Analysis in NK Cells from Patients Infected with Leishmania mexicana

    PubMed Central

    Castillo-Fernández, Juan E.; Miranda-Ortíz, Haydee; Fernández-López, Juan C.; Becker, Ingeborg; Rangel-Escareño, Claudia

    2016-01-01

    An important NK-cell inhibition with reduced TNF-α, IFN-γ and TLR2 expression had previously been identified in patients with diffuse cutaneous leishmaniasis (DCL) infected with Leishmania mexicana. In an attempt to pinpoint alterations in the signaling pathways responsible for the NK-cell dysfunction in patients with DCL, this study aimed at identifying differences in the NK-cell response towards Leishmania mexicana lipophosphoglycan (LPG) between patients with localized and diffuse cutaneous leishmaniasis through gene expression profiling. Our results indicate that important genes involved in the innate immune response to Leishmania are down-regulated in NK cells from DCL patients, particularly TLR and JAK/STAT signaling pathways. This down-regulation showed to be independent of LPG stimulation. The study sheds new light for understanding the mechanisms that undermine the correct effector functions of NK cells in patients with diffuse cutaneous leishmaniasis contributing to a better understanding of the pathobiology of leishmaniasis. PMID:27031998

  10. Application of hydrostatic CCC-TLC-HPLC-ESI-TOF-MS for the bioguided fractionation of anticholinesterase alkaloids from Argemone mexicana L. roots.

    PubMed

    Kukula-Koch, Wirginia; Mroczek, Tomasz

    2015-03-01

    A rapid hydrostatic counter-current chromatography-thin-layer chromatography-electrospray-ionization time-of-flight mass spectrometry (CCC-TLC-ESI-TOF-MS) technique was established for use in seeking potent anti-Alzheimer's drugs among the acethylcholinesterase inhibitors in Argemone mexicana L. underground parts, with no need to isolate components in pure form. The dichloromethane extract from the roots of Mexican prickly poppy that was most rich in secondary metabolites was subjected to hydrostatic-CCC-based fractionation in descending mode, using a biphasic system composed of petroleum ether-ethyl acetate-methanol-water at the ratio of 1.5:3:2.1:2 (v/v). The obtained fractions were analyzed in a TLC-based AChE-inhibition "Fast Blue B" test. All active components in the fractions, including berberine, protopine, chelerithrine, sanguinarine, coptisine, palmatine, magnoflorine, and galanthamine, were identified in a direct TLC-HPLC-ESI-TOF-MS assay with high accuracy. This is the first time galanthamine has been reported in the extract of Mexican prickly poppy and the first time it has been identified in any member of the Papaveraceae family, in the significant quantity of 0.77%. PMID:25618762

  11. NK cell activity differs between patients with localized and diffuse cutaneous leishmaniasis infected with Leishmania mexicana: a comparative study of TLRs and cytokines.

    PubMed

    Cañeda-Guzmán, Isabel Cristina; Salaiza-Suazo, Norma; Fernández-Figueroa, Edith A; Carrada-Figueroa, Georgina; Aguirre-García, Magdalena; Becker, Ingeborg

    2014-01-01

    Leishmania mexicana causes localized (LCL) or diffuse cutaneous leishmaniasis (DCL). The cause of dissemination in DCL remains unknown, yet NK cells possibly play a role in activating leishmanicidal mechanisms during innate and adaptive immune responses. We had previously shown that Leishmania lipophosphoglycan (LPG) is a ligand for TLR2, activating human NK cells. We have now analyzed NK cells in LCL and DCL patients. NK numbers and effector mechanisms differed drastically between both groups of patients: DCL patients showed reduced NK cell numbers; diminished IFN-γ and TNF-α production; and lower TLR2, TLR1, and TLR6 expression as compared to LCL patients. The altered protein expression found in NK cells of DCL patients correlated with their down-regulation of IFN-γ gene expression in LPG-stimulated and non-stimulated cells as compared to LCL patients. NK cell response was further analyzed according to gender, age, and disease evolution in LCL patients showing that female patients produced higher IFN-γ levels throughout the disease progression, whereas TLR2 expression diminished in both genders with prolonged disease evolution and age. We furthermore show the activation pathway of LPG binding to TLR2 and demonstrated that TLR2 forms immunocomplexes with TLR1 and TLR6. In addition to the reduced NK cell numbers in peripheral blood, DCL patients also showed reduced NK cell numbers in the lesions. They were randomly scattered within the lesions, showing diminished cytokine production, which contrasts with those of LCL lesions, where NK cells produced IFN-γ and TNF-α and were found within organized granulomas. We conclude that in DCL patients the reduced NK-cell numbers and their diminished activity, evidenced by low TLR expression and low cytokine production, are possibly involved in the severity of the disease. Our results provide new information on the contribution of NK cells in Leishmania infections of the human host. PMID:25397678

  12. Polycyclic aromatic hydrocarbons in urban tunnels of Guanajuato city (Mexico) measured in deposited dust particles and in transplanted lichen Xanthoparmelia mexicana (Gyeln.) Hale.

    PubMed

    Puy-Alquiza, María Jesús; Reyes, Veridiana; Wrobel, Katarzyna; Wrobel, Kazimierz; Torres Elguera, Julio César; Miranda-Aviles, Raúl

    2016-06-01

    Sixteen priority PAHs were determined in five urban tunnels of Guanajuato city, through which about 4 % of population walks and about 25,000 vehicles pass daily. Xanthoparmelia mexicana (Gyeln.) Hale, highly abundant lichen in this region, was exposed during 6 months and then the samples were collected together with the wall dust; both materials were analyzed by gas chromatography-mass spectrometry. Total PAH concentrations in dust and in lichen samples were in the range 1392-7961 ng g(-1) (average per tunnel 4637 ng g(-1)) and 522-3571 ng g(-1) (average 2587 ng g(-1)), respectively. In dust, the highest concentrations corresponded to PYR, FLA, BaA, CHR, BaP, and PHE, whereas in lichens the most abundant were DahA, IcdP, BghiP, and PYR. The obtained results suggested passive deposition of PAHs on lipophilic lichen surface rather than phenomena associated with metabolic activity of the exposed organisms. Application of seven different molecular diagnostic ratios pointed to gasoline-operated cars as the principal source of PAHs. Based on the obtained results and their comparison with data reported for other geographical regions, Guanajuato tunnels were considered moderately contaminated with PAHs; however toxic BaP equivalent concentrations integrated for seven carcinogenic compounds presented relatively high values in four tunnels: 567-1051 ngBaPeq g(-1) as evaluated for dust samples. Since up to 7000 persons walk daily through tunnels, the obtained data call for more detailed study evaluating PAHs toxicity in Guanajuato population. PMID:26961526

  13. Molecular Diversity between Salivary Proteins from New World and Old World Sand Flies with Emphasis on Bichromomyia olmeca, the Sand Fly Vector of Leishmania mexicana in Mesoamerica

    PubMed Central

    Townsend, Shannon; Pasos-Pinto, Silvia; Sanchez, Laura; Rasouli, Manoochehr; B. Guimaraes-Costa, Anderson; Aslan, Hamide; Francischetti, Ivo M. B.; Oliveira, Fabiano; Becker, Ingeborg; Kamhawi, Shaden; Ribeiro, Jose M. C.; Jochim, Ryan C.; Valenzuela, Jesus G.

    2016-01-01

    Background Sand fly saliva has been shown to have proteins with potent biological activities, salivary proteins that can be used as biomarkers of vector exposure, and salivary proteins that are candidate vaccines against different forms of leishmaniasis. Sand fly salivary gland transcriptomic approach has contributed significantly to the identification and characterization of many of these salivary proteins from important Leishmania vectors; however, sand fly vectors in some regions of the world are still neglected, as Bichromomyia olmeca (formerly known as Lutzomyia olmeca olmeca), a proven vector of Leishmania mexicana in Mexico and Central America. Despite the importance of this vector in transmitting Leishmania parasite in Mesoamerica there is no information on the repertoire of B. olmeca salivary proteins and their relationship to salivary proteins from other sand fly species. Methods and Findings A cDNA library of the salivary glands of wild-caught B. olmeca was constructed, sequenced, and analyzed. We identified transcripts encoding for novel salivary proteins from this sand fly species and performed a comparative analysis between B. olmeca salivary proteins and those from other sand fly species. With this new information we present an updated catalog of the salivary proteins specific to New World sand flies and salivary proteins common to all sand fly species. We also report in this work the anti-Factor Xa activity of Lofaxin, a salivary anticoagulant protein present in this sand fly species. Conclusions This study provides information on the first transcriptome of a sand fly from Mesoamerica and adds information to the limited repertoire of salivary transcriptomes from the Americas. This comparative analysis also shows a fast degree of evolution in salivary proteins from New World sand flies as compared with Old World sand flies. PMID:27409591

  14. Stage-Specific Adhesion of Leishmania Promastigotes to Sand Fly Midguts Assessed Using an Improved Comparative Binding Assay

    PubMed Central

    Wilson, Raymond; Bates, Michelle D.; Dostalova, Anna; Jecna, Lucie; Dillon, Rod J.; Volf, Petr; Bates, Paul A.

    2010-01-01

    Background The binding of Leishmania promastigotes to the midgut epithelium is regarded as an essential part of the life-cycle in the sand fly vector, enabling the parasites to persist beyond the initial blood meal phase and establish the infection. However, the precise nature of the promastigote stage(s) that mediate binding is not fully understood. Methodology/Principal Findings To address this issue we have developed an in vitro gut binding assay in which two promastigote populations are labelled with different fluorescent dyes and compete for binding to dissected sand fly midguts. Binding of procyclic, nectomonad, leptomonad and metacyclic promastigotes of Leishmania infantum and L. mexicana to the midguts of blood-fed, female Lutzomyia longipalpis was investigated. The results show that procyclic and metacyclic promastigotes do not bind to the midgut epithelium in significant numbers, whereas nectomonad and leptomonad promastigotes both bind strongly and in similar numbers. The assay was then used to compare the binding of a range of different parasite species (L. infantum, L. mexicana, L. braziliensis, L. major, L. tropica) to guts dissected from various sand flies (Lu. longipalpis, Phlebotomus papatasi, P. sergenti). The results of these comparisons were in many cases in line with expectations, the natural parasite binding most effectively to its natural vector, and no examples were found where a parasite was unable to bind to its natural vector. However, there were interesting exceptions: L. major and L. tropica being able to bind to Lu. longipalpis better than L. infantum; L. braziliensis was able to bind to P. papatasi as well as L. major; and significant binding of L. major to P. sergenti and L. tropica to P. papatasi was observed. Conclusions/Significance The results demonstrate that Leishmania gut binding is strictly stage-dependent, is a property of those forms found in the middle phase of development (nectomonad and leptomonad forms), but is absent

  15. New neotropical species of Opiinae (Hymenoptera, Braconidae) reared from fruit-infesting and leaf-mining Tephritidae (Diptera) with comments on the  Diachasmimorpha mexicana species group and the genera Lorenzopius and Tubiformopius

    PubMed Central

    Wharton, Robert; Ward, Lauren; Miko, Istvan

    2012-01-01

    Abstract Four new species of opiine Braconidae are described from Mexico. These are Diachasmimorpha martinalujai Wharton reared from Rhagoletis infesting fruits of Crataegus spp., Diachasmimorpha norrbomi Wharton reared from Euphranta mexicana infesting fruits of Ribes pringlei, Eurytenes (Stigmatopoea) norrbomi Wharton reared from Trypeta concolor mining leaves of Barkleyanthus salicifolia and Eurytenes (Stigmatopoea) maya Wharton reared from Rhagoletis pomonella infesting apples and fruits of Crataegus spp. Morphological features of the first metasomal segment and occipital carina, useful for placement of these species, are discussed relative to the genera Diachasmimorpha, Eurytenes, Lorenzopius, Tubiformopius, and Opius s.l. Descriptions and diagnoses are referenced to the Hymenoptera Anatomy Ontology. The following represent new combinations: Diachasmimorpha hildagensis, Lorenzopius euryteniformis, and Tubiformopius tubibasis. Revised diagnoses are provided for Diachasmimorpha hildagensis, Diachasmimorpha mexicana, Diachasmimorpha sanguinea, Eurytenes (Stigmatopoea), Lorenzopius, Lorenzopius euryteniformis, Tubiformopius, Tubiformopius tubigaster, Tubiformopius tubibasis, Opius incoligma, and Opius rugicoxis. Two species groups are delineated within Lorenzopius and a key to species of Diachasmimorpha occurring in the New World is provided. PMID:23818811

  16. O- and N-glycosylation of the Leishmania mexicana-secreted acid phosphatase. Characterization of a new class of phosphoserine-linked glycans.

    PubMed

    Ilg, T; Overath, P; Ferguson, M A; Rutherford, T; Campbell, D G; McConville, M J

    1994-09-30

    The protozoan parasite Leishmania mexicana secretes a heavily glycosylated 100-kDa acid phosphatase (sAP) which is associated with one or more polydisperse proteophosphoglycans. Most of the glycans in this complex were released using mild acid hydrolysis conditions that preferentially cleave phosphodiester linkages. The released saccharides were shown to consist of monomeric mannose and a series of neutral and phosphorylated glycans by Dionex high performance liquid chromatography, methylation analysis, exoglycosidase digestions, and one-dimensional 1H NMR spectroscopy. The neutral species comprised a linear series of oligosaccharides with the structures [Man alpha 1-2]1-5Man. The phosphorylated oligosaccharides were characterized as PO4-6Gal beta 1-4Man and PO4-6[Glc beta 1-3]Gal beta 1-4Man. The attachment of these glycans to the polypeptide backbone via the linkage, Man alpha 1-PO4-Ser, is suggested by: 1) the finding that more than 60% of the serine residues in the polypeptide are phosphorylated and 2) the resistance of the phosphoserine residues to alkaline phosphatase digestion unless the sAP was first treated with either mild acid (to release all glycans) or jack bean alpha-mannosidase (to release neutral mannose glycans). Analysis of the partially resolved components of the complex indicated that the most of the O-linked glycans on the 100-kDa phosphoglycoprotein comprised mannose and the mannose-oligosaccharides. In contrast the major O-linked glycans on the proteophosphoglycan were short phosphoglycan chains, containing on average two repeat units per chain. In addition to the O-linked glycans, both components in the sAP complex contained N-linked glycans. The N-glycanase F-released glycans were characterized by Bio-Gel P4 chromatography and exoglycosidase digestions to be the biantennary oligomannose type with the structures Glc1Man6GlcNAc2 and Man6GlcNAc2. The O-linked glycans of the sAP complex are similar to those found in the phosphoglycan chains of

  17. La Artesania Mexicana (Mexican Handicrafts).

    ERIC Educational Resources Information Center

    Steele, Bettina

    This booklet contains instructions in English and Spanish for making eleven typical Mexican craft articles. The instructions are accompanied by pen-and-ink drawings. The objects are (1) "La Rosa" (The Rose); (2) "El Crisantemo" (The Chrysanthemum); (3) "La Amapola" (The Poppy); (4) "Ojos de Dios" (God's Eyes); (5) "Ojitos con dos caras" (Two-Sided…

  18. A unique, highly conserved secretory invertase is differentially expressed by promastigote developmental forms of all species of the human pathogen, Leishmania

    PubMed Central

    Lyda, Todd A.; Joshi, Manju B.; Andersen, John F.; Kelada, Andrew Y.; Owings, Joshua P.; Bates, Paul A.; Dwyer, Dennis M.

    2015-01-01

    Leishmania are protozoan pathogens of humans that exist as extracellular promastigotes in the gut of their sand fly vectors and as obligate intracellular amastigotes within phagolysosomes of infected macrophages. Between infectious blood meal feeds, sand flies take plant juice meals that contain sucrose and store these sugars in their crop. Such sugars are regurgitated into the sand fly anterior midgut where they impact the developing promastigote parasite population. In this report we showed that promastigotes of all Leishmania species secreted an invertase/sucrase enzyme during their growth in vitro. In contrast, neither L. donovani nor L. mexicana amastigotes possessed any detectable invertase activity. Importantly, no released/secreted invertase activity was detected in culture supernatants from either Trypanosoma brucei or Trypanosoma cruzi. Using HPLC, the L. donovani secretory invertase was isolated and subjected to amino acid sequencing. Subsequently, we used a molecular approach to identify the LdINV and LmexINV genes encoding the ~72 kDa invertases produced by these organisms. Interestingly, we identified high fidelity LdINV-like homologs in the genomes of all Leishmania sp. but none were present in either T. brucei or T. cruzi. Northern blot and RT-PCR analyses showed that these genes were developmentally/differentially expressed in promastigotes but not amastigotes of these parasites. Homologous transfection studies demonstrated that these genes in fact encoded the functional secretory invertases produced by these parasites. Cumulatively, our results suggest that these secretory enzymes play critical roles in the survival/growth/development and transmission of all Leishmania parasites within their sand fly vector hosts. PMID:25763714

  19. A Mexicana Woman's Education in 1980s Chicago

    ERIC Educational Resources Information Center

    Rivera, Angelica

    2012-01-01

    The purpose of this paper is to describe the educational experiences of a Mexican woman in Chicago's Little Village community in the 1980s. The author was born in Mexico and raised in Little Village which has been home to Mexican immigrant communities for decades. Little Village is also home to the largest Mexican-American population in the…

  20. Proteome profiling of Leishmania infantum promastigotes.

    PubMed

    Alcolea, Pedro J; Alonso, Ana; Larraga, Vicente

    2011-01-01

    A proteome analysis of the promastigote stage of the trypanosomatid parasite Leishmania infantum (MON-1 zymodeme) is described here for the first time. Total protein extracts were prepared at early logarithmic and stationary phases of replicate axenic cultures and processed by 2D electrophoresis (pH 3-10). A total of 28 differentially regulated proteins were identified by matrix-assisted laser desorption/ionization-tandem time of flight mass spectrometry. This approach has revealed that the electron transfer flavoprotein (ETF) and the eukaryotic elongation factor 1α (eEF1α) subunit have the same differential expression pattern at the protein and mRNA levels, up-regulation in the stationary phase. A low-molecular-weight isoform and an alternatively processed form of the eEF1α subunit have been detected. A 51 kDa subunit of replication factor A is up-regulated in dividing logarithmic promastigotes. None of the proteins described here shows opposite differential regulation values with the corresponding mRNA levels. Taken together with previous approaches to the proteome and the transcriptome, this report contributes to the elucidation of the differential regulation patterns of the ETF, the eEF1α subunit, the 40S ribosomal protein S12, α-tubulin and the T-complex protein 1 subunit γ throughout the life cycle of the parasites from the genus Leishmania. PMID:21569158

  1. Attenuation of Leishmania infantum chagasi Metacyclic Promastigotes by Sterol Depletion

    PubMed Central

    Gaur Dixit, Upasna; Barker, Jason H.; Teesch, Lynn M.; Love-Homan, Laurie; Donelson, John E.; Wilson, Mary E.

    2013-01-01

    The infectious metacyclic promastigotes of Leishmania protozoa establish infection in a mammalian host after they are deposited into the dermis by a sand fly vector. Several Leishmania virulence factors promote infection, including the glycosylphosphatidylinositol membrane-anchored major surface protease (MSP). Metacyclic Leishmania infantum chagasi promastigotes were treated with methyl-beta-cyclodextrin (MβCD), a sterol-chelating reagent, causing a 3-fold reduction in total cellular sterols as well as enhancing MSP release without affecting parasite viability in vitro. MβCD-treated promastigotes were more susceptible to complement-mediated lysis than untreated controls and reduced the parasite load 3-fold when inoculated into BALB/c mice. Paradoxically, MβCD-treated promastigotes caused a higher initial in vitro infection rate in human or murine macrophages than untreated controls, although their intracellular multiplication was hindered upon infection establishment. There was a corresponding larger amount of covalently bound C3b than iC3b on the parasite surfaces of MβCD-treated promastigotes exposed to healthy human serum in vitro, as well as loss of MSP, a protease that enhances C3b cleavage to iC3b. Mass spectrometry showed that MβCD promotes the release of proteins into the extracellular medium, including both MSP and MSP-like protein (MLP), from virulent metacyclic promastigotes. These data support the hypothesis that plasma membrane sterols are important for the virulence of Leishmania protozoa at least in part through retention of membrane virulence proteins. PMID:23630964

  2. Antiprotozoal activity of Neurolaena lobata.

    PubMed

    Berger, I; Passreiter, C M; Cáceres, A; Kubelka, W

    2001-06-01

    Extracts, fractions and sesquiterpene lactones from Neurolaena lobata (L.) R. Br. (Asteraceae), a traditional medicinal plant from Guatemala, were tested in vitro against Leishmania spp. promastigotes, Trypanosoma cruzi trypomastigotes and epimastigotes and Trichomonas vaginalis trophozoites. The ethanol extract inhibited the parasite growth of L. mexicana, T. cruzi and T. vaginalis significantly. The pure germacranolides 1 and a mixture of 2 and 3, isolated from the ethonal extract, were highly active against L. mexicana and T. cruzi. PMID:11406857

  3. Tamoxifen Induces Apoptosis of Leishmania major Promastigotes in Vitro

    PubMed Central

    Doroodgar, Masoud; Delavari, Mahdi; Doroodgar, Moein; Abbasi, Ali; Taherian, Ali Akbar; Doroodgar, Abbas

    2016-01-01

    Tamoxifen is an antagonist of the estrogen receptor and currently used for the treatment of breast cancer. The current treatment of cutaneous leishmaniasis with pentavalent antimony compounds is not satisfactory. Therefore, in this study, due to its antileishmanial activity, effects of tamoxifen on the growth of promastigotes and amastigotes of Leishmania major Iranian strain were evaluated in vitro. Promastigotes and amastigotes were treated with different concentrations (1, 5, 10, 20, and 50 μg/ml) and time periods (24, 48, and 72 hr) of tamoxifen. After tamoxifen treatment, MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5 biphenyl tetrazolium bromide assay) was used to determine the percentage of live parasites and Graph Pad Prism software to calculate IC50. Flow cytometry was applied to investigate the induction of tamoxifen-induced apoptosis in promastigotes. The half maximal inhibitory concentration (IC50) of tamoxifen on promastigotes was 2.6 μg/ml after 24 hr treatment. Flow cytometry analysis showed that tamoxifen induced early and late apoptosis in Leishmania promastigotes. While after 48 hr in control group the apoptosis was 2.0%, the 50 µg/L concentration of tamoxifen increased it to 59.7%. Based on the in vitro antileishmanial effect, tamoxifen might be used for leishmaniasis treatment; however, further researches on in vivo effects of tamoxifen in animal models are needed. PMID:26951973

  4. Leishmania infantum: infection of macrophages in vitro with promastigotes.

    PubMed

    Méndez, S; Nell, M; Alunda, J M

    1996-06-01

    Leishmania infantum promastigotes in axenic culture exhibit limited infectivity for mouse peritoneal macrophages (M phi) in vitro using standard culture conditions (37 degrees C; 95% air/5% CO2) compared to Leishmania donovani promastigotes which induce notable infections. The infectivity of logarithmic (log) and stationary (stat) phase promastigotes of L. infantum was enhanced by the addition of fresh homologous serum, but no amastigotes were observed after 4 days. Prolonged infections, including transformation and survival of intracellular amastigotes in BALB/c mouse and hamster resident peritoneal M phi and M phi cell line J774.G8 were obtained by incubating M phi for 48 h at 26 degrees C prior to standard culture. Enhanced infectivity was observed in a number of L. infantum strains subject to this transient thermal change. PMID:8875306

  5. Leishmania promastigotes: building a safe niche within macrophages.

    PubMed

    Moradin, Neda; Descoteaux, Albert

    2012-01-01

    Upon their internalization by macrophages, Leishmania promastigotes inhibit phagolysosome biogenesis. The main factor responsible for this inhibition is the promastigote surface glycolipid lipophosphoglycan (LPG). This glycolipid has a profound impact on the phagosome, causing periphagosomal accumulation of F-actin and disruption of phagosomal lipid microdomains. Functionally, this LPG-mediated inhibition of phagosome maturation is characterized by an impaired assembly of the NADPH oxidase and the exclusion of the vesicular proton-ATPase from phagosomes. In this chapter, we review the current knowledge concerning the nature of the intra-macrophage compartment in which Leishmania donovani promastigotes establish infection. We also describe how LPG enables this parasite to remodel the parasitophorous vacuole. PMID:23050244

  6. Leishmania promastigotes: building a safe niche within macrophages

    PubMed Central

    Moradin, Neda; Descoteaux, Albert

    2012-01-01

    Upon their internalization by macrophages, Leishmania promastigotes inhibit phagolysosome biogenesis. The main factor responsible for this inhibition is the promastigote surface glycolipid lipophosphoglycan (LPG). This glycolipid has a profound impact on the phagosome, causing periphagosomal accumulation of F-actin and disruption of phagosomal lipid microdomains. Functionally, this LPG-mediated inhibition of phagosome maturation is characterized by an impaired assembly of the NADPH oxidase and the exclusion of the vesicular proton-ATPase from phagosomes. In this chapter, we review the current knowledge concerning the nature of the intra-macrophage compartment in which Leishmania donovani promastigotes establish infection. We also describe how LPG enables this parasite to remodel the parasitophorous vacuole. PMID:23050244

  7. The activity of ozonated olive oil against Leishmania major promastigotes

    PubMed Central

    Rajabi, Omid; Sazgarnia, Ameneh; Abbasi, Fatemeh; Layegh, Pouran

    2015-01-01

    Objective(s): Cutaneous Leishmaniasis is a common and endemic disease in Khorasan province in North-East of Iran. The pentavalant antimony (Sb V) is the mainstay of treatment that has many side effects and resistance to the drug has been reported. The microbicidal effect of ozone was proven in different microorganisms. Since there is no study in this respect and to achieve a low cost and effective treatment, we decided to evaluate the efficacy of ozone against promastigotes of Leishmania major, in vitro. Materials and Methods: Ozonated olive oil was prepared after production of ozone by bubbling ozone-oxygen gas produced by ozone generator through olive oil until it solidified. Promastigotes of L. major were cultivated in two phasic media. After calculation of the number of promastigotes, they were incubated with ozonated olive oil (0, 0.626, 0.938, 1.25, 2.5, 5, 10 mcg/ml) at 28 °c for 24 hr. Parasites survival percentage was evaluated using MTS and microscopic assay, and then compared with Glucantime and non-ozonated olive oil. Results: According to the results, there were significant differences in parasites survival percentage between ozonated olive oil and non-ozonated olive oil, at similar concentrations (P<0.001). Ozonated olive oil was more effective than Glucantime. According to MTS results, Glucantime and ozonated olive oil gel concentrations that are required to inhibit the growth of L. major promastigotes by 50% (IC50), were 165 and 0.002 mg/ml, respectively. Conclusion: Ozonated olive oil has in vitro activity against the promastigotes of L. major and this effect is dose dependent. PMID:26523224

  8. Activity of (-)alpha-bisabolol against Leishmania infantum promastigotes.

    PubMed

    Morales-Yuste, M; Morillas-Márquez, F; Martín-Sánchez, J; Valero-López, A; Navarro-Moll, M C

    2010-03-01

    Many of the drugs used to treat leishmaniasis are associated with numerous adverse effects. Agents of natural origin have shown activity against different parasites. With this background, an in vitro study was conducted on the activity of (-)alpha-bisabolol, the principal component of Chamomilla recutita essential oil, against Leishmania infantum promastigotes, the main species responsible for human leishmaniasis in Spain. At the two highest concentrations tested (1000 and 500mug/ml), (-)alpha-bisabolol and pentamidine (control agent) achieved 100% inhibition of L. infantum promastigote. These in vitro data can be considered promising in support of the therapeutic use of (-)alpha-bisabolol preparations to treat leishmaniasis caused by L. infantum species. PMID:19577452

  9. Destruction of Leishmania mexicana amazonensis amastigotes within macrophages by lysosomotropic amino acid esters

    PubMed Central

    1986-01-01

    Leishmania amastigotes parasitize almost exclusively the mononuclear phagocytes of mammals. The organisms survive and multiply within acidified vacuoles (parasitophorous vacuoles; p.v.) akin to phagolysosomes. Certain amino acid esters are known to accumulate in and disrupt lysosomes. We postulated that, since Leishmania possess lysosome-like organelles, they may be susceptible to the potentially high ester concentrations attained in the p.v. We report here that L- amino acid esters can rapidly destroy intracellular Leishmania at concentrations that do not appear to damage the host cells. L-leu-OMe, which cured greater than or equal to 90% of infected macrophages at 0.8 mM concentrations, was used in most of the experiments. L-leu-OMe was only active after infection, implying inefficient transfer from secondary lysosomes to the p.v. Parasite destruction had several features in common with lysosomal and leukocyte damage induced by the esters, i.e., inactivity of D-amino acid esters, a marked pH dependence and increased killing after ester pulses at lower temperatures. Killing depended on the amino acid and on the ester substitution. The most active of the methyl esters assayed was that of leucine, followed by those of tryptophan, glutamic acid, methionine, phenylalanine, and tyrosine. Methyl esters of seven other amino acids were inactive when tested at up to 10 mM concentrations. Among leucine esters studied, benzyl ester was sixfold more active than the methyl homolog. The dipeptide L-leu-leu-OMe produced 90% cure at 0.08 mM concentrations. Leishmanicidal activity could be related to penetration of the parasites by the esters or to toxic ester hydrolysis products released in the p.v. The first hypothesis is supported by the pH-dependent destruction of isolated amastigotes by the esters. Furthermore, relatively high concentrations of L-leucine, methanol, or benzyl alcohol were not demonstrably toxic to the amastigotes. We postulate that ester concentrations sufficient to damage the intracellular amastigotes may be obtained within the p.v. after exposure of infected macrophages to the esters. Esters preferentially hydrolyzed by parasite enzymes may be expected to be leishmanicidal, but less damaging to the host. PMID:3950542

  10. Unidad: Las influencias culturales en el arte mexicana (Unit: Cultural Influences in Mexican Art). Dos semanas.

    ERIC Educational Resources Information Center

    Finere, Neal

    This two-week unit, appropriate for bilingual education settings as well as foreign language programs, deals with the three primary cultural influences found in Mexican contemporary art. The multisensory materials, pragmatic focus, and direct creative student involvement are designed to make it a microcosmic, real-life experience. The first part…

  11. Verbascoside Inhibits Promastigote Growth and Arginase Activity of Leishmania amazonensis.

    PubMed

    Maquiaveli, Claudia C; Lucon-Júnior, João F; Brogi, Simone; Campiani, Giuseppe; Gemma, Sandra; Vieira, Paulo C; Silva, Edson R

    2016-05-27

    Verbascoside (1) is a phenylethanoid glycoside that has antileishmanial activity against Leishmania infantum and Leishmania donovani. In this study, we verified the activity of 1 on Leishmania amazonensis and arginase inhibition. Compound 1 showed an EC50 of 19 μM against L. amazonensis promastigotes and is a competitive arginase inhibitor (Ki = 0.7 μM). Docking studies were performed to assess the interaction of 1 with arginase at the molecular level. Arginase is an enzyme of the polyamine biosynthesis pathway that is important to parasite infectivity, and the results of our study suggest that 1 could be useful to develop new approaches for treating leishmaniasis. PMID:27096224

  12. Influence of the Microenvironment in the Transcriptome of Leishmania infantum Promastigotes: Sand Fly versus Culture

    PubMed Central

    Alcolea, Pedro J.; Alonso, Ana; Domínguez, Mercedes; Parro, Víctor; Jiménez, Maribel; Molina, Ricardo; Larraga, Vicente

    2016-01-01

    Zoonotic visceral leishmaniasis is a vector-borne disease caused by Leishmania infantum in the Mediterranean Basin, where domestic dogs and wild canids are the main reservoirs. The promastigote stage replicates and develops within the gut of blood-sucking phlebotomine sand flies. Mature promastigotes are injected in the dermis of the mammalian host and differentiate into the amastigote stage within parasitophorous vacuoles of phagocytic cells. The major vector of L. infantum in Spain is Phlebotomus perniciosus. Promastigotes are routinely axenized and cultured to mimic in vitro the conditions inside the insect gut, which allows for most molecular, cellular, immunological and therapeutical studies otherwise inviable. Culture passages are known to decrease infectivity, which is restored by passage through laboratory animals. The most appropriate source of promastigotes is the gut of the vector host but isolation of the parasite is technically challenging. In fact, this option is not viable unless small samples are sufficient for downstream applications like promastigote cultures and nucleic acid amplification. In this study, in vitro infectivity and differential gene expression have been studied in cultured promastigotes at the stationary phase and in promastigotes isolated from the stomodeal valve of the sand fly P. perniciosus. About 20 ng RNA per sample could be isolated. Each sample contained L. infantum promastigotes from 20 sand flies. RNA was successfully amplified and processed for shotgun genome microarray hybridization analysis. Most differentially regulated genes are involved in regulation of gene expression, intracellular signaling, amino acid metabolism and biosynthesis of surface molecules. Interestingly, meta-analysis by hierarchical clustering supports that up-regulation of 22.4% of the differentially regulated genes is specifically enhanced by the microenvironment (i.e. sand fly gut or culture). The correlation between cultured and naturally

  13. Influence of the Microenvironment in the Transcriptome of Leishmania infantum Promastigotes: Sand Fly versus Culture.

    PubMed

    Alcolea, Pedro J; Alonso, Ana; Domínguez, Mercedes; Parro, Víctor; Jiménez, Maribel; Molina, Ricardo; Larraga, Vicente

    2016-05-01

    Zoonotic visceral leishmaniasis is a vector-borne disease caused by Leishmania infantum in the Mediterranean Basin, where domestic dogs and wild canids are the main reservoirs. The promastigote stage replicates and develops within the gut of blood-sucking phlebotomine sand flies. Mature promastigotes are injected in the dermis of the mammalian host and differentiate into the amastigote stage within parasitophorous vacuoles of phagocytic cells. The major vector of L. infantum in Spain is Phlebotomus perniciosus. Promastigotes are routinely axenized and cultured to mimic in vitro the conditions inside the insect gut, which allows for most molecular, cellular, immunological and therapeutical studies otherwise inviable. Culture passages are known to decrease infectivity, which is restored by passage through laboratory animals. The most appropriate source of promastigotes is the gut of the vector host but isolation of the parasite is technically challenging. In fact, this option is not viable unless small samples are sufficient for downstream applications like promastigote cultures and nucleic acid amplification. In this study, in vitro infectivity and differential gene expression have been studied in cultured promastigotes at the stationary phase and in promastigotes isolated from the stomodeal valve of the sand fly P. perniciosus. About 20 ng RNA per sample could be isolated. Each sample contained L. infantum promastigotes from 20 sand flies. RNA was successfully amplified and processed for shotgun genome microarray hybridization analysis. Most differentially regulated genes are involved in regulation of gene expression, intracellular signaling, amino acid metabolism and biosynthesis of surface molecules. Interestingly, meta-analysis by hierarchical clustering supports that up-regulation of 22.4% of the differentially regulated genes is specifically enhanced by the microenvironment (i.e. sand fly gut or culture). The correlation between cultured and naturally

  14. Differences in human macrophage receptor usage, lysosomal fusion kinetics and survival between logarithmic and metacyclic Leishmania infantum chagasi promastigotes

    PubMed Central

    Ueno, Norikiyo; Bratt, Carol L.; Rodriguez, Nilda E.; Wilson, Mary E.

    2009-01-01

    Summary The obligate intracellular protozoan, Leishmania infantum chagasi (Lic) undergoes receptor-mediated phagocytosis by macrophages, followed by a transient delay in phagolysosome maturation. We found differences in the pathway through which virulent Lic metacyclic promastigotes or avirulent logarithmic promastigotes are phagocytosed by human monocyte-derived macrophages (MDMs). Both logarithmic and metacyclic promastigotes entered MDMs through a compartment lined by the third complement receptor (CR3). In contrast, many logarithmic promastigotes entered through vacuoles lined by mannose receptors (MR) whereas most metacyclic promastigotes did not (p < 0.005). CR3 positive vacuoles containing metacyclic promastigotes stained for caveolin-1 protein, suggesting CR3 localizes in caveolae during phagocytosis. Following entry, the kinetics of phagolysosomal maturation and intracellular survival also differed. Vacuoles containing metacyclic parasites did not accumulate lysosome-associated membrane protein-1 (LAMP-1) at early times after phagocytosis, whereas vacuoles with logarithmic promastigotes did. MDMs phagocytosed greater numbers of logarithmic than metacyclic promastigotes, yet metacyclics ultimately replicated intracellularly with greater efficiency. These data suggest that virulent metacyclic Leishmania promastigotes fail to ligate macrophage MR, and enter through a path that ultimately enhances intracellular survival. The relatively quiescent entry of virulent Leishmania spp. into macrophages may be accounted for by the ability of metacyclic promastigotes to selectively bypass deleterious entry pathways. PMID:19702651

  15. Proteomic analysis of antigens from Leishmania infantum promastigotes.

    PubMed

    Dea-Ayuela, María Auxiliadora; Rama-Iñiguez, Sara; Bolás-Fernández, Francisco

    2006-07-01

    Leishmaniasis is a zoonotic disease caused by the species of the genus Leishmania, flagellated protozoa that multiply inside mammalian macrophages and are transmitted by the bite of the sandfly. The disease is widespread and due to the lack of fully effective treatment and vaccination the search for new drugs and immune targets is needed. Proteomics seems to be a suitable strategy because the annotated sequenced genome of L. major is available. Here, we present a high-resolution proteome for L. infantum promastigotes comprising of around 700 spots. Western blot with rabbit hyperimmune serum raised against L. infantum promastiogote extracts and further analysis by MALDI-TOF and MALDI-TOF/TOF MS allowed the identification of various relevant functional antigenic proteins. Major antigenic proteins were identified as propionil carboxilasa, ATPase beta subunit, transketolase, proteasome subunit, succinyl-diaminopimelate desuccinylase, a probable tubulin alpha chain, the full-size heat shock protein 70, and several proteins of unknown function. In addition, one enzyme from the ergosterol biosynthesis pathway (adrenodoxin reductase) and the structural paraflagellar rod protein 3 (PAR3) were found among non-antigenic proteins. This study corroborates the usefulness of proteomics in identifying new proteins with crucial biological functions in Leishmania parasites. PMID:16791830

  16. Hydrogen peroxide-mediated toxicity for Leishmania donovani chagasi promastigotes. Role of hydroxyl radical and protection by heat shock.

    PubMed Central

    Zarley, J H; Britigan, B E; Wilson, M E

    1991-01-01

    Leishmania must survive despite exposure to the toxic oxidant hydrogen peroxide (H2O2) during phagocytosis by macrophages. We investigated the mechanism of H2O2 toxicity for L. donovani chagasi promastigotes, and factors responsible for their relative H2O2 resistance. There was a dose-dependent toxic effect of H2O2 for promastigotes isolated during logarithmic phase of growth. In contrast, stationary phase promastigotes were less susceptible to H2O2 toxicity, and more infectious for BALB/c mice. By spin trapping we found that hydroxyl radical (.OH) was generated after exposure of promastigotes to H2O2, and the amount of .OH was greater with log-phase than with stationary-phase promastigotes. .OH was generated after the addition of H2O2 to the cytosol but not the membranes of fractionated promastigotes, and the magnitude of .OH was greater in log than in stationary promastigote cytosol. Deferoxamine inhibition suggested that intracellular promastigote iron catalyzes .OH formation via the Fenton reaction. Furthermore, exposure of log-phase promastigotes to heat shock induced a relative H2O2-resistant state, which was not associated with a decrease in .OH formation but which required ongoing transcription. Thus, growth to stationary phase and heat shock both induce a state of relative H2O2 resistance, but these are probably due to different resistance mechanisms. Images PMID:1658042

  17. Leishmania Promastigotes Lack Phosphatidylserine but Bind Annexin V upon Permeabilization or Miltefosine Treatment

    PubMed Central

    Zampieri, Ricardo Andrade; Gonzaga dos Santos, Marcos; Schiller, Jürgen; Pomorski, Thomas Günther

    2012-01-01

    The protozoan parasite Leishmania is an intracellular pathogen infecting and replicating inside vertebrate host macrophages. A recent model suggests that promastigote and amastigote forms of the parasite mimic mammalian apoptotic cells by exposing phosphatidylserine (PS) at the cell surface to trigger their phagocytic uptake into host macrophages. PS presentation at the cell surface is typically analyzed using fluorescence-labeled annexin V. Here we show that Leishmania promastigotes can be stained by fluorescence-labeled annexin V upon permeabilization or miltefosine treatment. However, combined lipid analysis by thin-layer chromatography, mass spectrometry and 31P nuclear magnetic resonance (NMR) spectroscopy revealed that Leishmania promastigotes lack any detectable amount of PS. Instead, we identified several other phospholipid classes such phosphatidic acid, phosphatidylethanolamine; phosphatidylglycerol and phosphatidylinositol as candidate lipids enabling annexin V staining. PMID:22870283

  18. Cryptolepine-Induced Cell Death of Leishmania donovani Promastigotes Is Augmented by Inhibition of Autophagy

    PubMed Central

    Sengupta, Souvik; Chowdhury, Sayan; BoseDasgupta, Somdeb; Wright, Colin W.; Majumder, Hemanta K.

    2011-01-01

    Leishmania donovani are the causative agents of visceral leishmaniasis worldwide. Lack of vaccines and emergence of drug resistance warrants the need for improved drug therapy and newer therapeutic intervention strategies against leishmaniasis. In the present study, we have investigated the effect of the natural indoloquinoline alkaloid cryptolepine on L. donovani AG83 promastigotes. Our results show that cryptolepine induces cellular dysfunction in L. donovani promastigotes, which leads to the death of this unicellular parasite. Interestingly, our study suggest that cryptolepine-induced cell death of L. donovani is counteracted by initial autophagic features elicited by the cells. For the first time, we show that autophagy serves as a survival mechanism in response to cryptolepine treatment in L. donovani promastigotes and inhibition of autophagy causes an early increase in the amount of cell death. This study can be exploited for designing better drugs and better therapeutic strategies against leishmaniasis in future. PMID:22091398

  19. Leishmania donovani lacking the Golgi GDP-Man transporter LPG2 exhibit attenuated virulence in mammalian hosts.

    PubMed

    Gaur, Upasna; Showalter, Melissa; Hickerson, Suzanne; Dalvi, Rahul; Turco, Salvatore J; Wilson, Mary E; Beverley, Stephen M

    2009-07-01

    Surface phosophoglycans such as lipophosphoglycan (LPG) or proteophosphoglycan (PPG) and glycosylinositol phospholipids (GIPLs) modulate essential interactions between Leishmania and mammalian macrophages. Phosphoglycan synthesis depends on the Golgi GDP-mannose transporter encoded by LPG2. LPG2-null (lpg2(-)) Leishmania major cannot establish macrophage infections or induce acute pathology, whereas lpg2(-)Leishmania mexicana retain virulence. lpg2(-)Leishmania donovani has been reported to survive poorly in cultured macrophages but in vivo survival has not been explored. Herein we discovered that, similar to lpg2(-)L. major, lpg2(-)L. donovani promastigotes exhibited diminished virulence in mice, but persisted at consistently low levels. lpg2(-)L. donovani promastigotes could not establish infection in macrophages and could not transiently inhibit phagolysosomal fusion. Furthermore, lpg2(-) promastigotes of L. major, L. donovani and L. mexicana were highly susceptible to complement-mediated lysis. We conclude that phosphoglycan assembly and expression mediated by L. donovani LPG2 are important for promastigote and amastigote virulence, unlike L. mexicana but similar to L. major. PMID:19328787

  20. Leishmania donovani lacking the Golgi GDP-Man transporter LPG2 exhibit attenuated virulence in mammalian hosts

    PubMed Central

    Gaur, Upasna; Showalter, Melissa; Hickerson, Suzanne; Dalvi, Rahul; Turco, Salvatore J.; Wilson, Mary E.; Beverley, Stephen M.

    2009-01-01

    Surface phosophoglycans such as lipophosphoglycan (LPG) or proteophosphoglycan (PPG) and glycosylinositol phospholipids (GIPLs) modulate essential interactions between Leishmania and mammalian macrophages. Phosphoglycan synthesis depends on the Golgi GDP-mannose transporter encoded by LPG2. LPG2-null (lpg2−) L. major cannot establish macrophage infections or induce acute pathology, whereas lpg2− L. mexicana retain virulence. lpg2− L. donovani has been reported to survive poorly in cultured macrophages but in vivo survival has not been explored. Herein we discovered that, similar to lpg2− L. major, lpg2− L. donovani promastigotes exhibited diminished virulence in mice, but persisted at consistently low levels. lpg2− L. donovani promastigotes could not establish infections macrophages and could not transiently inhibit phagolysosomal fusion. Furthermore, lpg2− promastigotes of L. major, L. donovani and L. mexicana were highly susceptible to complement mediated lysis. We conclude that phosphoglycan assembly and expression mediated by L. donovani LPG2 are important for promastigote and amastigote virulence, unlike L. mexicana but similar to L. major. PMID:19328787

  1. Los Sentimientos, Luchas, y Esperanzas de Cinco Familias Mexicana: The Sentiments, Struggles, and Hopes of Five Mexican Families

    ERIC Educational Resources Information Center

    Dueñas, Gilbert

    2015-01-01

    Within the everyday space of their modest trailer homes, I observed compelling environments in the households of 5 Mexican families. In their homes, there existed "los padres con sus hijos" (parents with their children) who intuitively expressed "cariño," or love, for one another, conversed in multiple languages, practiced…

  2. Our Treasures: A Celebration of Nebraska's Mexican Heritage = Nuestros Tesoros: Una celebracion de la herencia mexicana de Nebraska.

    ERIC Educational Resources Information Center

    Nebraska State Historical Society, Lincoln.

    This publication is intended as a resource guide for teachers, students, and librarians interested in Mexican American culture, and for groups wishing to conduct local community research using oral histories. The Mexican American Traditions in Nebraska project was designed to document and preserve the richness of Nebraska's Mexican culture. The…

  3. Cell differentiation and infectivity of Leishmania mexicana are inhibited in a strain resistant to an ABC-transporter blocker.

    PubMed

    Silva, N; Camacho, N; Figarella, K; Ponte-Sucre, A

    2004-06-01

    We analysed whether markers of cell differentiation and infectivity differed when compared to the parental sensitive strain [NR(Gs)] in an in vitro selected Leishmania strain [NR(Gr)] resistant to Glibenclamide, an ATP-binding-cassette (ABC)-transporter blocker. The data show that the cell body area was larger in NR(Gr) compared to NR(Gs) and that functional characters associated with an infective metacyclic phenotype, such as resistance to the lytic effect of the alternative complement pathway and expression of the Meta-1 protein, were reduced. The infectivity of NR(Gr) to J774.1 macrophages was also significantly reduced. These results suggest that resistance in Leishmania against Glibenclamide, a general blocker of P-glycoproteins, could produce functional modifications that may be relevant for Leishmania differentiation, infectivity and survival. PMID:15206465

  4. The triterpenoid fraction from Trichosanthes dioica root exhibits in vitro antileishmanial effect against Leishmania donovani promastigotes

    PubMed Central

    Bhattacharya, Sanjib; Biswas, Moulisha; Haldar, Pallab K.

    2013-01-01

    Background: Trichosanthes dioica Roxb. (Cucurbitaceae), called pointed gourd in English is a dioecious climber found wild throughout the plains of the Indian subcontinent and traditionally used in India for several medicinal purposes. Objective: The present study was aimed at the evaluation of in vitro antileishmanial effect of triterpenoid fraction from T. dioica root (CETD). Materials and Methods: The antileishmanial activity of CETD was evaluated against Leishmania donovani (strain MHOM/IN/83/AG83)) promastigotes by in vitro promastigote cell toxicity assay by using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide). Potassium antimonyl tartrate was used as reference. Results: Here, CETD markedly inhibited the growth of L. donovani promastigotes in vitro in a concentration dependent manner and demonstrated IC50 value of 18.75 μg/ml. The reference drug potassium antimonyl tartrate exhibited IC50 of 7.52 μg/ml. Conclusion: From the present study it can be inferred that the triterpenoid fraction of T. dioica root exhibited remarkable antileishmanial activity against Leishmania donovani promastigotes in vitro. PMID:23798885

  5. Effects of nitro-heterocyclic derivatives against Leishmania (Leishmania) infantum promastigotes and intracellular amastigotes.

    PubMed

    Petri e Silva, Simone Carolina Soares; Palace-Berl, Fanny; Tavares, Leoberto Costa; Soares, Sandra Regina Castro; Lindoso, José Angelo Lauletta

    2016-04-01

    Leishmaniasis is an overlooked tropical disease affecting approximately 1 million people in several countries. Clinical manifestation depends on the interaction between Leishmania and the host's immune response. Currently available treatment options for leishmaniasis are limited and induce severe side effects. In this research, we tested nitro-heterocyclic compounds (BSF series) as a new alternative against Leishmania. Its activity was measured in Leishmania (Leishmania) infantum promastigotes and intracellular amastigotes using MTT colorimetric assay. Additionally, we assessed the phosphatidylserine exposure by promastigotes, measured by flow cytometry, as well as nitric oxide production, measured by Griess' method. The nitro-heterocyclic compounds (BSF series) showed activity against L. (L.) infantum promastigotes, inducting the phosphatidylserine exposition by promastigotes, decreasing intracellular amastigotes and increasing oxide nitric production. The selectivity index was more prominent to Leishmania than to macrophages. Compared to amphotericin b, our compounds presented higher IC50, however the selectivity index was more specific to parasite than to amphotericin b. In conclusion, these nitro-heterocyclic compounds showed to be promising as an anti-Leishmania drug, in in vitro studies. PMID:26795261

  6. Leishmanicidal evaluation of extracts from native plants of the Yucatan peninsula.

    PubMed

    Peraza-Sánchez, S R; Cen-Pacheco, F; Noh-Chimal, A; May-Pat, F; Simá-Polanco, P; Dumonteil, E; García-Miss, M R; Mut-Martín, M

    2007-06-01

    Methanol extracts were prepared from different parts of 18 plants collected in the Yucatan peninsula and evaluated in an in vitro bioassay for leishmanicidal activity against Leishmania mexicana promastigotes. The ten most potent plant extracts (IC(50)<50 microg/ml) were Aphelandra scabra leaves, Byrsonima bucidaefolia bark, Byrsonima crassifolia bark, Clusia flava leaves, Cupania dentata bark, Diphysa carthagenensis leaves, Dorstenia contrajerva whole plant, Milleria quinqueflora roots, Tridax procumbens whole plant, and Vitex gaumeri bark. PMID:17482379

  7. Selection and phenotype characterisation of sitamaquine-resistant promastigotes of Leishmania donovani.

    PubMed

    Bories, Christian; Cojean, Sandrine; Huteau, Françoise; Loiseau, Philippe M

    2008-03-01

    A Leishmania donovani promastigote line resistant to 160microM sitamaquine, named SITA-160R, was selected in vitro by continuous stepwise drug pressure. SITA-160R line was able to infect murine peritoneal macrophages in vitro in the same manner as the wild-type line (WT) but was less infective for Balb/c mice than its parent WT clone. This line was about five and three times more resistant to sitamaquine than the WT line on promastigote and intramacrophage amastigote forms, respectively. No cross-resistance with other antileishmanial agents was observed and this resistance was stable when parasites were subcultured in drug-free medium for a long time or after in vivo passage. PMID:18249083

  8. Comparative real-time kinetic analysis of human complement killing of Leishmania infantum promastigotes derived from axenic culture or from Phlebotomus perniciosus.

    PubMed

    Moreno, Inmaculada; Molina, Ricardo; Toraño, Alfredo; Laurin, Edurne; García, Esther; Domínguez, Mercedes

    2007-01-01

    Although Leishmania metacyclic promastigotes are generally considered resistant to human complement, studies of in vitro-cultured axenic stationary promastigotes using serum concentrations that approximate physiological plasma conditions indicate complement sensitivity. Natural Leishmania infection is caused by sand fly-inoculated promastigotes, whose complement resistance has not been analyzed systematically. We compared Leishmania susceptibility to human complement in L. infantum promastigotes derived from in vitro cultures and from sand flies. Phlebotomus perniciosus sand flies were fed with axenic promastigotes, L. infantum-infected U-937 cells, or spleen cells from L. infantum-infected hamsters. On selected days post-feeding, flies were dissected and promastigotes isolated; in addition, axenic promastigotes were obtained from culture at equivalent days of growth. In near-physiological serum concentration and temperature conditions, measurement of real-time kinetics of propidium iodide uptake showed that approximately 90% of axenic- and sand fly-derived promastigotes were rapidly killed by complement. We found no substantial differences between promastigotes from axenic culture, those isolated from flies on different post-feeding days, or those generated in flies fed with distinct inocula. The results indicate that Leishmania susceptibility to human complement is independent of promastigote developmental stage in the sand fly mid-gut and in axenic culture. PMID:18023393

  9. Identification of a macrophage-binding determinant on lipophosphoglycan from Leishmania major promastigotes.

    PubMed

    Kelleher, M; Bacic, A; Handman, E

    1992-01-01

    Leishmania are obligatory intracellular parasites in mammalian macrophages that gain entry by receptor-mediated phagocytosis. Their major cell surface glycoconjugate, lipophosphoglycan (LPG), has been implicated in this process. A monoclonal antibody specific for Leishmania major LPG (WIC 79.3), which has been shown to block promastigote attachment to macrophages, was used to identify a macrophage-binding determinant of LPG. WIC 79.3 bound exclusively to the phosphorylated repeats of LPG and not to the saccharide core or lipid anchor. Furthermore, the epitope recognized by WIC 79.3 mapped to the phosphorylated oligosaccharide P5b, PO4-6[Gal(beta 1-3)Gal(beta 1-3)Gal(beta 1-3)]Gal(beta 1-4)Man(alpha 1-, which is unique to the LPG of promastigotes of L.major. Phosphorylated oligosaccharides P3, PO4-6[Gal(beta 1-3)[Gal(beta 1-4) Man(alpha 1-, and P4b, PO4-6[Gal(beta 1-3)Gal(beta 1-3)] Gal(beta 1-4)Man(alpha 1-, were also recognized by WIC 79.3 but with considerably lower (approximately 100-fold) affinities. The phosphorylated oligosaccharide P5b inhibited attachment of promastigotes of L. major to the macrophage cell line J774 to the same degree as phosphoglycan (derived from LPG) and Fab fragments of WIC 79.3, suggesting that P5b is a site of L. major LPG that is recognized by macrophage receptor(s) and is an important determinant in the attachment of promastigotes to host macrophages and initiation of infection. PMID:1370357

  10. Effect of Aqueous Allium cepa and Ixora brachiata Root Extract on Leishmania major Promastigotes

    PubMed Central

    Sadeghi-Nejad, Batool; Saki, Jasem

    2014-01-01

    Background: Leishmaniasis is a major worldwide public health problem with about two to three million humans threatened by this disease annually. Allium cepa (onion) is an important dietary vegetable and was used as a herbal medicine for centuries. The root of Ixora brachiata is medicinally important. Objectives: The aim of this study was to investigate the anti-Leishmania effect of the ethanolic and methanolic extracts of Ixora brachiata root and aqueous onion extracts on Leishmania major promastigotes. Patients and Methods: The parasites isolated from cutaneous leishmaniasis were exposed with different concentrations of selected plant extracts and their inhibitory effects on the promastigotes were evaluated after 24 and 48 hours. Results: Among tested plant extracts, Ixora brachiata root extracts revealed the best activity against Leishmania major promastigotes with IC100 value of 2.5 mg/mL and IC50 value of 0.078 mg/mL. Conclusions: This study showed that aqueous Allium cepa and Ixora brachiata root extracts as natural products could be used as alternative drugs in treatment of leishmaniasis. PMID:24872942

  11. Ultrastructural Analysis of Leishmania infantum chagasi Promastigotes Forms Treated In Vitro with Usnic Acid

    PubMed Central

    da Luz, João S. B.; de Oliveira, Erwelly B.; Martins, Monica C. B.; da Silva, Nicácio H.; Alves, Luiz C.; dos Santos, Fábio A. B.; da Silva, Luiz L. S.; Silva, Eliete C.; de Medeiros, Paloma L.

    2015-01-01

    Leishmaniasis is considered by the World Health Organization as one of the infectious parasitic diseases endemic of great relevance and a global public health problem. Pentavalent antimonials used for treatment of this disease are limited and new phytochemicals emerge as an alternative to existing treatments, due to the low toxicity and cost reduction. Usnic acid is uniquely found in lichens and is especially abundant in genera such as Alectoria, Cladonia, Evernia, Lecanora, Ramalina, and Usnea. Usnic acid has been shown to exhibit antiviral, antiprotozoal, antiproliferative, anti-inflammatory, and analgesic activity. The aim of this study was to evaluate the antileishmanial activity of usnic acid on Leishmania infantum chagasi promastigotes and the occurrence of drug-induced ultrastructural damage in the parasite. Usnic acid was effective against the promastigote forms (IC50 = 18.30 ± 2.00 µg/mL). Structural and ultrastructural aspects of parasite were analyzed. Morphological alterations were observed as blebs in cell membrane and shapes given off, increasing the number of cytoplasmic vacuoles, and cellular and mitochondrial swelling, with loss of cell polarity. We concluded that the usnic acid presented antileishmanial activity against promastigote forms of Leishmania infantum chagasi and structural and ultrastructural analysis reinforces its cytotoxicity. Further, in vitro studies are warranted to further evaluate this potential. PMID:25767824

  12. Leishmania major Promastigotes Evade LC3-Associated Phagocytosis through the Action of GP63

    PubMed Central

    Matte, Christine; Casgrain, Pierre-André; Séguin, Olivier; Moradin, Neda; Hong, Wan Jin; Descoteaux, Albert

    2016-01-01

    The protozoan Leishmania parasitizes macrophages and evades the microbicidal consequences of phagocytosis through the inhibition of phagolysosome biogenesis. In this study, we investigated the impact of this parasite on LC3-associated phagocytosis, a non-canonical autophagic process that enhances phagosome maturation and functions. We show that whereas internalization of L. major promastigotes by macrophages promoted LC3 lipidation, recruitment of LC3 to phagosomes was inhibited through the action of the parasite surface metalloprotease GP63. Reactive oxygen species generated by the NOX2 NADPH oxidase are necessary for LC3-associated phagocytosis. We found that L. major promastigotes prevented, in a GP63-dependent manner, the recruitment of NOX2 to phagosomes through a mechanism that does not involve NOX2 cleavage. Moreover, we found that the SNARE protein VAMP8, which regulates phagosomal assembly of the NADPH oxidase NOX2, was down-modulated by GP63. In the absence of VAMP8, recruitment of LC3 to phagosomes containing GP63-deficient parasites was inhibited, indicating that VAMP8 is involved in the phagosomal recruitment of LC3. These findings reveal a role for VAMP8 in LC3-associated phagocytosis and highlight a novel mechanism exploited by L. major promastigotes to interfere with the host antimicrobial machinery. PMID:27280768

  13. Leishmania major Promastigotes Evade LC3-Associated Phagocytosis through the Action of GP63.

    PubMed

    Matte, Christine; Casgrain, Pierre-André; Séguin, Olivier; Moradin, Neda; Hong, Wan Jin; Descoteaux, Albert

    2016-06-01

    The protozoan Leishmania parasitizes macrophages and evades the microbicidal consequences of phagocytosis through the inhibition of phagolysosome biogenesis. In this study, we investigated the impact of this parasite on LC3-associated phagocytosis, a non-canonical autophagic process that enhances phagosome maturation and functions. We show that whereas internalization of L. major promastigotes by macrophages promoted LC3 lipidation, recruitment of LC3 to phagosomes was inhibited through the action of the parasite surface metalloprotease GP63. Reactive oxygen species generated by the NOX2 NADPH oxidase are necessary for LC3-associated phagocytosis. We found that L. major promastigotes prevented, in a GP63-dependent manner, the recruitment of NOX2 to phagosomes through a mechanism that does not involve NOX2 cleavage. Moreover, we found that the SNARE protein VAMP8, which regulates phagosomal assembly of the NADPH oxidase NOX2, was down-modulated by GP63. In the absence of VAMP8, recruitment of LC3 to phagosomes containing GP63-deficient parasites was inhibited, indicating that VAMP8 is involved in the phagosomal recruitment of LC3. These findings reveal a role for VAMP8 in LC3-associated phagocytosis and highlight a novel mechanism exploited by L. major promastigotes to interfere with the host antimicrobial machinery. PMID:27280768

  14. Developmental modification of lipophosphoglycan during the differentiation of Leishmania major promastigotes to an infectious stage.

    PubMed

    McConville, M J; Turco, S J; Ferguson, M A; Sacks, D L

    1992-10-01

    Protozoan parasites of the genus Leishmania produce the novel surface glycoconjugate, lipophosphoglycan (LPG), which is required for parasite infectivity. In this study we show that LPG structure is modified during the differentiation of L. major promastigotes from a less infectious form in logarithmic growth phase to a highly infectious 'metacyclic' form during stationary growth phase. In both stages, the LPGs comprise linear chains of phosphorylated oligosaccharide repeat units which are anchored to the membrane via a glycosyl-phosphatidylinositol glycolipid anchor. During metacyclogenesis there is (i) an approximate doubling in the average number of repeat units per molecule from 14 to 30, (ii) a pronounced decrease in the relative abundance of repeat units with side chains of beta Gal or Gal beta 1-3Gal beta 1-, and a corresponding increase in repeat units with either no side chains or with side chains of Arap alpha 1-2 Gal beta 1- and (iii) a decrease in the frequency with which the glycolipid anchor is substituted with a single glucose alpha 1-phosphate residue. While the majority of the LPG phosphoglycan chains are capped with the neutral disaccharide, Man alpha 1-2Man, a significant minority of the chains appeared to terminate in non-phosphorylated repeat units and may represent incompletely capped species. We suggest that the developmental modification of LPG may be important in modulating the binding of promastigotes to receptors in the sandfly midgut and on human macrophages and in increasing the resistance of metacyclic promastigotes to complement-mediated lysis. PMID:1396559

  15. In vitro activity of sulfonamides and sulfones against Leishmania major promastigotes.

    PubMed Central

    Peixoto, M P; Beverley, S M

    1987-01-01

    We examined the susceptibility of promastigotes of Leishmania major to sulfonamides and sulfones in vitro. In a completely defined medium only sulfamoxole, sulfaquinoxaline, and dapsone were inhibitory; the concentrations required for 50% inhibition of the rate of growth were 150, 600, and 600 microM, respectively. Eleven other sulfa drugs were ineffective at concentrations up to 2 mM. The growth inhibition was similar to that observed in procaryotes: the cells continued logarithmic growth for several cell doublings before inhibition was observed. Surprisingly, the addition of p-aminobenzoate or folate did not reverse the effects of the active sulfa drugs, the effects of sulfamoxole and methotrexate were additive rather than synergistic, and the addition of thymidine reversed methotrexate but not sulfa-drug inhibition. These results suggest that the mode of action of sulfa drugs on L. major is not by the classical route of inhibition of de novo folate synthesis. Promastigotes could be propagated for more than 40 passages in a completely defined medium in which the only added pterin was biopterin. The folate concentration in this medium was less than 10(-10) to 10(-11) M, as determined by a Leishmania bioassay. Although these data suggest that L. major may be capable of de novo synthesis of folate, the nonclassical mode of action of sulfa drugs, as well as other studies, favors the view that L. major is auxotrophic for folate. PMID:3435106

  16. Identification of Proteins in Promastigote and Amastigote-like Leishmania Using an Immunoproteomic Approach

    PubMed Central

    Coelho, Vinicio T. S.; Oliveira, Jamil S.; Valadares, Diogo G.; Chávez-Fumagalli, Miguel A.; Duarte, Mariana C.; Lage, Paula S.; Soto, Manuel; Santoro, Marcelo M.; Tavares, Carlos A. P.; Fernandes, Ana Paula; Coelho, Eduardo A. F.

    2012-01-01

    Background The present study aims to identify antigens in protein extracts of promastigote and amastigote-like Leishmania (Leishmania) chagasi syn. L. (L.) infantum recognized by antibodies present in the sera of dogs with asymptomatic and symptomatic visceral leishmaniasis (VL). Methodology/Principal Findings Proteins recognized by sera samples were separated by two-dimensional electrophoresis (2DE) and identified by mass spectrometry. A total of 550 spots were observed in the 2DE gels, and approximately 104 proteins were identified. Several stage-specific proteins could be identified by either or both classes of sera, including, as expected, previously known proteins identified as diagnosis, virulence factors, drug targets, or vaccine candidates. Three, seven, and five hypothetical proteins could be identified in promastigote antigenic extracts; while two, eleven, and three hypothetical proteins could be identified in amastigote-like antigenic extracts by asymptomatic and symptomatic sera, as well as a combination of both, respectively. Conclusions/Significance The present study represents a significant contribution not only in identifying stage-specific L. infantum molecules, but also in revealing the expression of a large number of hypothetical proteins. Moreover, when combined, the identified proteins constitute a significant source of information for the improvement of diagnostic tools and/or vaccine development to VL. PMID:22272364

  17. Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani Promastigotes

    PubMed Central

    Singh, Arpita; Mandal, Debjani

    2016-01-01

    The promastigote stage of Leishmania resides in the sand fly gut, enriched with sugar molecules. Recently we reported that Leishmania donovani possesses a sucrose uptake system and a stable pool of intracellular sucrose metabolizing enzyme. In the present study, we purified the intracellular sucrase nearly to its homogeneity and compared it with the purified extracellular sucrase. The estimated size of intracellular sucrase is ~112 kDa by gel filtration chromatography, native PAGE, and substrate staining. However, in SDS-PAGE, the protein is resolved at ~56 kDa, indicating the possibility of a homodimer in its native state. The kinetics of purified intracellular sucrase shows its higher substrate affinity with a K m of 1.61 mM than the extracellular form having a K m of 4.4 mM. The highly specific activity of intracellular sucrase towards sucrose is optimal at pH 6.0 and at 30°C. In this report the purification and characterization of intracellular sucrase provide evidence that sucrase enzyme exists at least in two different forms in Leishmania donovani promastigotes. This intracellular sucrase may support further intracellular utilization of transported sucrose. PMID:27190649

  18. In vitro leishmanicidal activities of sesquiterpene lactones from Tithonia diversifolia against Leishmania braziliensis promastigotes and amastigotes.

    PubMed

    de Toledo, Juliano S; Ambrósio, Sergio R; Borges, Carly H G; Manfrim, Viviane; Cerri, Daniel G; Cruz, Angela K; Da Costa, Fernando B

    2014-01-01

    Natural compounds represent a rich and promising source of novel, biologically active chemical entities for treating leishmaniasis. Sesquiterpene lactones are a recognized class of terpenoids with a wide spectrum of biological activities, including activity against Leishmania spp. In this work, a sesquiterpene lactone-rich preparation-a leaf rinse extract (LRE) from Tithonia diversifolia-was tested against promastigote forms of L. braziliensis. The results revealed that the LRE is a rich source of potent leishmanicidal compounds, with an LD50 value 1.5 ± 0.50 µg·mL-1. Therefore, eight sesquiterpene lactones from the LRE were initially investigated against promastigote forms of L. braziliensis. One of them did not present any significant leishmanicidal effect (LD50 > 50 µg·mL-1). Another had a cytotoxic effect against macrophages (4.5 µg·mL-1). The five leishmanicidal compounds with the highest level of selectivity were further evaluated against intracellular parasites (amastigotes) using peritoneal macrophages. Tirotundin 3-O-methyl ether, tagitinin F, and a guaianolide reduced the internalization of parasites after 48 h, in comparison with the negative control. This is the first report on sesquiterpene lactones that have potent leishmanicidal effects on both developmental stages of L. braziliensis. PMID:24830711

  19. Leishmania amazonensis promastigotes in 3D Collagen I culture: an in vitro physiological environment for the study of extracellular matrix and host cell interactions

    PubMed Central

    Rodrigues, Juliany C.F.; Viana, Nathan B.; Pontes, Bruno; Pereira, Camila F.A.; Silva-Filho, Fernando C.

    2014-01-01

    Leishmania amazonensis is the causative agent of American cutaneous leishmaniasis, an important neglected tropical disease. Once Leishmania amazonensis is inoculated into the human host, promastigotes are exposed to the extracellular matrix (ECM) of the dermis. However, little is known about the interaction between the ECM and Leishmania promastigotes. In this study we established L. amazonensis promastigote culture in a three-dimensional (3D) environment mainly composed of Collagen I (COL I). This 3D culture recreates in vitro some aspects of the human host infection site, enabling the study of the interaction mechanisms of L. amazonensis with the host ECM. Promastigotes exhibited “freeze and run” migration in the 3D COL I matrix, which is completely different from the conventional in vitro swimming mode of migration. Moreover, L. amazonensis promastigotes were able to invade, migrate inside, and remodel the 3D COL I matrix. Promastigote trans-matrix invasion and the freeze and run migration mode were also observed when macrophages were present in the matrix. At least two classes of proteases, metallo- and cysteine proteases, are involved in the 3D COL I matrix degradation caused by Leishmania. Treatment with a mixture of protease inhibitors significantly reduced promastigote invasion and migration through this matrix. Together our results demonstrate that L. amazonensis promastigotes release proteases and actively remodel their 3D environment, facilitating their migration. This raises the possibility that promastigotes actively interact with their 3D environment during the search for their cellular “home”—macrophages. Supporting this hypothesis, promastigotes migrated faster than macrophages in a novel 3D co-culture model. PMID:24765565

  20. In vivo and in vitro phagocytosis of Leishmania (Leishmania) amazonensis promastigotes by B-1 cells.

    PubMed

    Geraldo, M M; Costa, C R; Barbosa, F M C; Vivanco, B C; Gonzaga, W F K M; Novaes E Brito, R R; Popi, A F; Lopes, J D; Xander, P

    2016-06-01

    Leishmaniasis is caused by Leishmania parasites that infect several cell types. The promastigote stage of Leishmania is internalized by phagocytic cells and transformed into the obligate intracellular amastigote form. B-1 cells are a subpopulation of B cells that are able to differentiate in vitro and in vivo into mononuclear phagocyte-like cells with phagocytic properties. B-1 cells use several receptors for phagocytosis, such as the mannose receptor and third complement receptor. Leishmania binds to the same receptors on macrophages. In this study, we demonstrated that phagocytes derived from B-1 cells (B-1 CDP) were able to internalize promastigotes of L. (L.) amazonensis in vitro. The internalized promastigotes differentiated into amastigotes. Our results showed that the phagocytic index was higher in B-1 CDP compared to peritoneal macrophages and bone marrow-derived macrophages. The in vivo phagocytic ability of B-1 cells was also demonstrated. Parasites were detected inside purified B-1 cells after intraperitoneal infection with L. (L.) amazonensis promastigotes. Intraperitoneal stimulation with the parasites led to an increase in both IL-10 and TNF-α. These results highlight the importance of studying B-1 CDP cells as phagocytic cells that can participate and contribute to immunity to parasites. PMID:27084328

  1. The Calpain Inhibitor MDL28170 Induces the Expression of Apoptotic Markers in Leishmania amazonensis Promastigotes

    PubMed Central

    Marinho, Fernanda A.; Gonçalves, Keyla C. S.; Oliveira, Simone S. C.; Gonçalves, Diego S.; Matteoli, Filipe P.; Seabra, Sergio H.; Oliveira, Ana Carolina S.; Bellio, Maria; Oliveira, Selma S.; Souto-Padrón, Thaïs; d'Avila-Levy, Claudia M.; Santos, André L. S.; Branquinha, Marta H.

    2014-01-01

    Background Human cutaneous leishmaniasis is caused by distinct species, including Leishmania amazonensis. Treatment of cutaneous leishmaniasis is far from satisfactory due to increases in drug resistance and relapses, and toxicity of compounds to the host. As a consequence for this situation, the development of new leishmanicidal drugs and the search of new targets in the parasite biology are important goals. Methodology/Principal Findings In this study, we investigated the mechanism of death pathway induced by the calpain inhibitor MDL28170 on Leishmania amazonensis promastigote forms. The combined use of different techniques was applied to contemplate this goal. MDL28170 treatment with IC50 (15 µM) and two times the IC50 doses induced loss of parasite viability, as verified by resazurin assay, as well as depolarization of the mitochondrial membrane, which was quantified by JC-1 staining. Scanning and transmission electron microscopic images revealed drastic alterations on the parasite morphology, some of them resembling apoptotic-like death, including cell shrinking, surface membrane blebs and altered chromatin condensation pattern. The lipid rearrangement of the plasma membrane was detected by Annexin-V labeling. The inhibitor also induced a significant increase in the proportion of cells in the sub-G0/G1 phase, as quantified by propidium iodide staining, as well as genomic DNA fragmentation, detected by TUNEL assay. In cells treated with MDL28170 at two times the IC50 dose, it was also possible to observe an oligonucleossomal DNA fragmentation by agarose gel electrophoresis. Conclusions/Significance The data presented in the current study suggest that MDL28170 induces apoptotic marker expression in promastigotes of L. amazonensis. Altogether, the results described in the present work not only provide a rationale for further exploration of the mechanism of action of calpain inhibitors against trypanosomatids, but may also widen the investigation of the

  2. Evaluation of Macroalgae Sulfated Polysaccharides on the Leishmania (L.) amazonensis Promastigote

    PubMed Central

    Pires, Camila Lehnhardt; Rodrigues, Selma Dzimidas; Bristot, Daniel; Gaeta, Henrique Hessel; de Oliveira Toyama, Daniela; Farias, Wladimir Ronald Lobo; Toyama, Marcos Hikari

    2013-01-01

    The sulfated polysaccharides from Solieria filiformis (Sf), Botryocladia occidentalis (Bo), Caulerpa racemosa (Cr) and Gracilaria caudata (Gc) were extracted and extensively purified. These compounds were then subjected to in vitro assays to evaluate the inhibition of these polysaccharides on the growth of Leishmania (L.) amazonensis promastigotes. Under the same assay conditions, only three of the four sulfated polysaccharides were active against L. amazonensis, and the polysaccharide purified from Cr was the most potent (EC50 value: 34.5 μg/mL). The polysaccharides derived from Bo and Sf demonstrated moderate anti-leishmanial activity (EC50 values of 63.7 μg/mL and 137.4 μg/mL). In addition, we also performed in vitro cytotoxic assays toward peritoneal macrophages and J774 macrophages. For the in vitro cytotoxicity assay employing J774 cells, all of the sulfated polysaccharides decreased cell survival, with CC50 values of 27.3 μg/mL, 49.3 μg/mL, 73.2 μg/mL, and 99.8 μg/mL for Bo, Cr, Gc, and Sf, respectively. However, none of the sulfated polysaccharides reduced the cell growth rate of the peritoneal macrophages. These results suggest that macroalgae contain compounds with various chemical properties that can control specific pathogens. According to our results, the assayed sulfated polysaccharides were able to modulate the growth rate and cell survival of Leishmania (L.) amazonensis promastigotes in in vitro assays, and these effects involved the interaction of the sulfated polysaccharides on the cell membrane of the parasites. PMID:23519148

  3. CrATP interferes in the promastigote-macrophage interaction in Leishmania amazonensis infection.

    PubMed

    Ennes-Vidal, V; Castro, R O S; Britto, C; Barrabin, H; D'Avila-Levy, C M; Moreira, O C

    2011-07-01

    Recent have shown the relationship between Ecto-Nucleoside-Triphosphate-Diphosphohydrolases (Ecto-NTPDases or ecto-nucleotidases) and virulence and infectivity in trypanosomatids. In this work, the inhibition of the ecto-ATPase activities and promastigote growth of Leishmania amazonensis by CrATP was characterized. Furthermore, this compound was used to investigate the role of ecto-nucleotidase in the interaction of L. amazonensis with resident peritoneal macrophages obtained from BALB/c mice. CrATP partially inhibits the ecto-ATPase activity, presenting Ki values of 575·7±199·1 and 383·5±79·0 μm, in the presence or absence of 5 mm MgCl2, respectively. The apparent Kms for ATP (2·9±0·5 mm to Mg2+-dependent ecto-ATPase and 0·4±0·2 mm to Mg2+-independent ecto-ATPase activities) are not significantly altered by CrATP, suggesting a reversible non-competitive inhibition of both enzymes. When CrATP was added to the cultivation medium at 500 μm, it drastically inhibited the cellular growth. The interaction of promastigote forms of L. amazonensis with BALB/c peritoneal macrophages is strongly affected by CrATP. When the parasites were treated with 500 μm CrATP before interacting with macrophages, the adhesion and endocytic indices were strongly reduced to 53·0±14·8% and 39·8±1·1%, respectively. These results indicate that ecto-nucleotidase plays an important role in the infection process caused by Leishmania amazonensis. PMID:21679488

  4. Leishmania major-human macrophage interactions: cooperation between Mac-1 (CD11b/CD18) and complement receptor type 1 (CD35) in promastigote adhesion.

    PubMed Central

    Rosenthal, L A; Sutterwala, F S; Kehrli, M E; Mosser, D M

    1996-01-01

    It has been suggested that the developmental maturation of Leishmania major promastigotes can affect their interaction with human complement receptors. To study this, we measured the adhesion of metacyclic and logarithmic-phase L. major promastigotes to complement receptors expressed on primary macrophages, to recombinant receptors expressed on transfected cells, or to purified complement receptors in a cell-free system. We demonstrate that complement-opsonized promastigotes can bind to both Mac-1 and complement receptor type 1 (CR1) and that the transition of promastigotes from the noninfectious logarithmic phase of growth to the infectious metacyclic stage does not affect this interaction. Furthermore, we show that Mac-1 and CR1 can cooperate to mediate the efficient adhesion of complement-opsonized metacyclic promastigotes to cells expressing both receptors. On human monocyte-derived macrophages, Mac-1 appears to make a quantitatively greater contribution to this adhesion than does CR1, since blocking macrophage Mac-1 diminishes metacyclic promastigote adhesion to a greater extent than does blocking CR1. In addition, bovine monocytes lacking Mac-1 exhibit a dramatic decrease in complement-dependent promastigote adhesion, relative to normal monocytes. The predominance of Mac-1 in these interactions is due, at least in part, to the factor I cofactor activity of CR1, which facilitates the conversion of C3b to iC3b. The stable adhesion of complement-opsonized metacyclic promastigotes to Mac-1 is a prerequisite for phagocytosis by human monocyte-derived macrophages. Blocking Mac-1 on macrophages abrogates the majority of the complement-dependent phagocytosis of promastigotes, whereas blocking CR1 has no detectable effect on phagocytosis. In addition, bovine monocytes lacking Mac-1 exhibit a dramatic reduction in promastigote phagocytosis relative to normal bovine monocytes. We conclude, therefore, that the two complement receptors, Mac-1 and CR1, can cooperate to

  5. A family of heat shock protein 70-related genes are expressed in the promastigotes of Leishmania major.

    PubMed Central

    Searle, S; Campos, A J; Coulson, R M; Spithill, T W; Smith, D F

    1989-01-01

    We describe the isolation and characterisation of two novel genes of the parasitic protozoan Leishmania major that are related by nucleotide sequence homology to eukaryotic genes encoding 70 Kd. heat shock proteins. The transcription of neither gene is heat-inducible but both are constituitively-expressed throughout the promastigote stage of the parasite life cycle. A third gene shows differential expression between non-infective and infective promastigote stages in the absence of any temperature change. These genes are related by sequence homology to the tandemly-repeated hsp70 genes of trypanosomatids, but are located on different, dispersed chromosomes within the genome of L. major. The open reading frame for translation derived from one of these sequences contains a putative mitochondrial signal peptide at its amino-terminus. Images PMID:2762121

  6. Ultrastructural Changes and Death of Leishmania infantum Promastigotes Induced by Morinda citrifolia Linn. Fruit (Noni) Juice Treatment.

    PubMed

    Almeida-Souza, Fernando; Taniwaki, Noemi Nosomi; Amaral, Ana Cláudia Fernandes; de Souza, Celeste da Silva Freitas; Calabrese, Kátia da Silva; Abreu-Silva, Ana Lúcia

    2016-01-01

    The search for new treatments against leishmaniasis has increased due to high frequency of drug resistance registered in endemics areas, side effects, and complications caused by coinfection with HIV. Morinda citrifolia Linn., commonly known as Noni, has a rich chemical composition and various therapeutic effects have been described in the literature. Studies have shown the leishmanicidal activity of M. citrifolia; however, its action on the parasite has not yet been elucidated. In this work, we analyzed leishmanicidal activity and ultrastructural changes in Leishmania infantum promastigotes caused by M. citrifolia fruit juice treatment. M. citrifolia fruit extract showed a yield of 6.31% and high performance liquid chromatography identified phenolic and aromatic compounds as the major constituents. IC50 values were 260.5 µg/mL for promastigotes and 201.3 µg/mL for intracellular amastigotes of L. infantum treated with M. citrifolia. Cytotoxicity assay with J774.G8 macrophages showed that M. citrifolia fruit juice was not toxic up to 2 mg/mL. Transmission electron microscopy showed cytoplasmic vacuolization, lipid inclusion, increased exocytosis activity, and autophagosome-like vesicles in L. infantum promastigotes treated with M. citrifolia fruit juice. M. citrifolia fruit juice was active against L. infantum in the in vitro model used here causing ultrastructural changes and has a future potential for treatment against leishmaniasis. PMID:27313649

  7. In vitro selection of miltefosine resistance in promastigotes of Leishmania donovani from Nepal: genomic and metabolomic characterization.

    PubMed

    Shaw, C D; Lonchamp, J; Downing, T; Imamura, H; Freeman, T M; Cotton, J A; Sanders, M; Blackburn, G; Dujardin, J C; Rijal, S; Khanal, B; Illingworth, C J R; Coombs, G H; Carter, K C

    2016-03-01

    In this study, we followed the genomic, lipidomic and metabolomic changes associated with the selection of miltefosine (MIL) resistance in two clinically derived Leishmania donovani strains with different inherent resistance to antimonial drugs (antimony sensitive strain Sb-S; and antimony resistant Sb-R). MIL-R was easily induced in both strains using the promastigote-stage, but a significant increase in MIL-R in the intracellular amastigote compared to the corresponding wild-type did not occur until promastigotes had adapted to 12.2 μM MIL. A variety of common and strain-specific genetic changes were discovered in MIL-adapted parasites, including deletions at the LdMT transporter gene, single-base mutations and changes in somy. The most obvious lipid changes in MIL-R promastigotes occurred to phosphatidylcholines and lysophosphatidylcholines and results indicate that the Kennedy pathway is involved in MIL resistance. The inherent Sb resistance of the parasite had an impact on the changes that occurred in MIL-R parasites, with more genetic changes occurring in Sb-R compared with Sb-S parasites. Initial interpretation of the changes identified in this study does not support synergies with Sb-R in the mechanisms of MIL resistance, though this requires an enhanced understanding of the parasite's biochemical pathways and how they are genetically regulated to be verified fully. PMID:26713880

  8. Ultrastructural Changes and Death of Leishmania infantum Promastigotes Induced by Morinda citrifolia Linn. Fruit (Noni) Juice Treatment

    PubMed Central

    Almeida-Souza, Fernando; Taniwaki, Noemi Nosomi; Amaral, Ana Cláudia Fernandes; de Souza, Celeste da Silva Freitas; Calabrese, Kátia da Silva; Abreu-Silva, Ana Lúcia

    2016-01-01

    The search for new treatments against leishmaniasis has increased due to high frequency of drug resistance registered in endemics areas, side effects, and complications caused by coinfection with HIV. Morinda citrifolia Linn., commonly known as Noni, has a rich chemical composition and various therapeutic effects have been described in the literature. Studies have shown the leishmanicidal activity of M. citrifolia; however, its action on the parasite has not yet been elucidated. In this work, we analyzed leishmanicidal activity and ultrastructural changes in Leishmania infantum promastigotes caused by M. citrifolia fruit juice treatment. M. citrifolia fruit extract showed a yield of 6.31% and high performance liquid chromatography identified phenolic and aromatic compounds as the major constituents. IC50 values were 260.5 µg/mL for promastigotes and 201.3 µg/mL for intracellular amastigotes of L. infantum treated with M. citrifolia. Cytotoxicity assay with J774.G8 macrophages showed that M. citrifolia fruit juice was not toxic up to 2 mg/mL. Transmission electron microscopy showed cytoplasmic vacuolization, lipid inclusion, increased exocytosis activity, and autophagosome-like vesicles in L. infantum promastigotes treated with M. citrifolia fruit juice. M. citrifolia fruit juice was active against L. infantum in the in vitro model used here causing ultrastructural changes and has a future potential for treatment against leishmaniasis. PMID:27313649

  9. Cytotoxic Screening and In Vitro Evaluation of Pentadecane Against Leishmania infantum Promastigotes and Amastigotes.

    PubMed

    Bruno, Federica; Castelli, Germano; Migliazzo, Antonella; Piazza, Maria; Galante, Antonella; Lo Verde, Vincenza; Calderone, Simona; Nucatolo, Giuseppa; Vitale, Fabrizio

    2015-12-01

    Pentadecane is an organic compound that is made up of primarily carbon and hydrogen atoms. Pentadecane is a floral volatile found in many different plants and essential oils and also in crude extracts of some plants, and it shows antimicrobial activity. This study investigated in vitro effects of pentadecane in Leishmania infantum parasites and found that it decreases growth by 86 ± 2% both in promastigotes (half maximal inhibitory concentration [IC50] = 65.3 μM) and in amastigotes (IC50 = 60.5 μM), resulting in a reduction of macrophage infection; growth inhibition was 77% at 300 μM. Analysis of propidium iodide incorporation in L. infantum , treated with pentadecane at 48 hr, suggested that cells were arresting in the sub-G0/G1 and G1 phases of the cell cycle, whereas cytotoxicity assay of pentadecane in immortalized cells lines DH82 and U937 and in primary epithelial cells of Cercopiteco showed that it caused negligible cytotoxic effect. This study shows that pentadecane has antimicrobial activity against L. infantum parasites in in vitro culture. PMID:26321352

  10. Modulation of macrophage mannose receptor affects the uptake of virulent and avirulent Leishmania donovani promastigotes.

    PubMed

    Chakraborty, P; Ghosh, D; Basu, M K

    2001-10-01

    The effect of oxidants and the anti-inflammatory steroid dexamethasone on the attachment and internalization of virulent and avirulent Leishmania donovani promastigotes by the macrophage mannosyl fucosyl receptor was examined. Oxidants and dexamethasone are known to down- and upregulate the expression of the mannose receptor. Macrophages, when treated with 500 microM H2O2 at 37 C for 30 min, stimulate about 45% inhibition in uptake of an avirulent strain (UR6), and 30 and 25% inhibition for virulent strains AG-83 and GE-I, respectively. Treatment of macrophages with dexamethasone for 20 hr resulted in a stimulation in uptake of the parasite. When UR6 was used, a 3-fold increase in uptake was observed compared with the controls. Parasite uptake was also inhibited by the H2O2-generating system, glucose/glucose oxidase; inhibition was blocked by catalase. Treatment of macrophages either with H2O2 or dexamethasone did not affect the binding of the advanced glycosylation end product-bovine serum albumin (AGE-BSA), the ligand for AGE receptor of macrophages. Similarly, indirect evidence also shows that both types 1 and 3 complement receptors (CR1, CR3) are not affected by these treatments, indicating that, besides the mannosyl fucosyl receptor, other receptors are minimally altered in the identified condition. These results suggest that the up- and downregulation of the mannose receptor of macrophages may play a role in affecting L. donovani infection. PMID:11695359

  11. Effect of human urine on cell cycle and infectivity of Leismania species promastigotes in vitro.

    PubMed

    Allahverdiyev, Adil M; Bagirova, Melahat; Elcicek, Serhat; Koc, Rabia Cakir; Oztel, Olga N

    2011-10-01

    In vitro cultivation of Leishmania parasites plays an important role in diagnosis and treatment of leishmaniasis and in vaccine and drug development studies. Conversely, long-term cultivation of Leishmania parasites usually results in decreased infectivity potential. Some studies reported a stimulatory effect of human urine in Leishmania promastigotes. However, there is no information about the effects of urine within culture on the infectivity of Leishmania parasites. Analysis of the effect of urine have showed that proliferation indexes were significantly increased in culture medium supplemented with human urine (L. tropica = 38.17 ± 5.12, L. donovani = 34.74 ± 5.6, L. major = 34.22 ± 4.66, and L. infantum 35.88 ± 6.40) than in controls. Infection indexes were 13 ± 1.7 for L. tropica, 55 ± 2.2 for L. infantum, 41 ± 3.14 for L. donovani, and 49 ± 3.26 for L. major. Our results showed that human urine increased the infectivity and proliferation of Leishmania parasites. PMID:21976564

  12. Effect of Human Urine on Cell Cycle and Infectivity of Leismania Species Promastigotes In Vitro

    PubMed Central

    Allahverdiyev, Adil M.; Bagirova, Melahat; Elcicek, Serhat; Koc, Rabia Cakir; Oztel, Olga N.

    2011-01-01

    In vitro cultivation of Leishmania parasites plays an important role in diagnosis and treatment of leishmaniasis and in vaccine and drug development studies. Conversely, long-term cultivation of Leishmania parasites usually results in decreased infectivity potential. Some studies reported a stimulatory effect of human urine in Leishmania promastigotes. However, there is no information about the effects of urine within culture on the infectivity of Leishmania parasites. Analysis of the effect of urine have showed that proliferation indexes were significantly increased in culture medium supplemented with human urine (L. tropica = 38.17 ± 5.12, L. donovani = 34.74 ± 5.6, L. major = 34.22 ± 4.66, and L. infantum 35.88 ± 6.40) than in controls. Infection indexes were 13 ± 1.7 for L. tropica, 55 ± 2.2 for L. infantum, 41 ± 3.14 for L. donovani, and 49 ± 3.26 for L. major. Our results showed that human urine increased the infectivity and proliferation of Leishmania parasites. PMID:21976564

  13. C5 induces different cell death pathways in promastigotes of Leishmania amazonensis.

    PubMed

    Mendes, Edevi Arbonelli; Desoti, Vânia Cristina; Silva, Sueli de Oliveira; Ueda-Nakamura, Tânia; Dias Filho, Benedito Prado; Yamada-Ogatta, Sueli Fumie; Sarragiotto, Maria Helena; Nakamura, Celso Vataru

    2016-08-25

    Leishmaniasis is a neglected infection that is caused by Leishmania protozoa, affecting millions of people worldwide, mainly in tropical and subtropical regions. This disease has different clinical forms: cutaneous, mucocutaneous, and visceral. The drugs that are currently available for the treatment of this infection have limitations, such as high toxicity, long-term treatment, and leads to drug-resistant strains. Numerous studies, in various experimental models, have sought to develop more effective and less toxic chemotherapeutic agents against leishmaniasis. In the present study, we evaluated the mechanism of cell death that is induced by n-benzyl 1-(4-methoxy)phenyl-9H-β-carboline-3-carboxamide (C5) against Leishmania amazonensis. C5 increased reactive oxygen species production, depolarization of the mitochondrial membrane, DNA fragmentation, decrease of cell volume, lipoperoxidation, the accumulation of lipid bodies, and acidic vesicular organelles (AVOs) and caused the intense formation of autophagic compartments in L. amazonensis promastigotes. The results indicate that C5 causes L. amazonensis death through different pathways. PMID:27317947

  14. Golgi-mediated post-translational processing of secretory acid phosphatase by Leishmania donovani promastigotes.

    PubMed

    Bates, P A; Hermes, I; Dwyer, D M

    1990-03-01

    Monensin, an inhibitor of Golgi function, was used to investigate the role of this cell compartment in the glycosylation of Leishmania donovani promastigote secretory acid phosphatase (EC 3.1.3.2). Monensin-treated cells demonstrated morphological changes in the Golgi complex and secreted enzyme with an altered electrophoretic mobility: two discrete bands of approximately 95 and 110 kDa were found, as compared to the heterodisperse nature of the enzyme from untreated controls. Chemical deglycosylation by mild acid hydrolysis resulted in a similar effect on the electrophoretic mobility of purified extracellular enzyme. Acid phosphatase was also treated with N-glycosidase F (EC 3.5.1.52) to remove N-linked oligosaccharides. The altered lectin-binding properties of the enzyme after these two treatments demonstrated that an unusual type of galactose-containing acid-labile carbohydrate was present in secretory acid phosphatase in addition to the N-linked oligosaccharides. Further, experiments with 32P-labelled enzyme indicated that phosphodiester bonds were the structural component responsible for the sensitivity of this carbohydrate to mild acid hydrolysis. Cumulatively, these results demonstrated that a novel form of Golgi-mediated posttranslational modification had occurred to the secretory acid phosphatase presumably by the addition of an acid-labile phosphoglycan. PMID:2320058

  15. Leishmania amazonensis promastigotes induce and are killed by neutrophil extracellular traps

    PubMed Central

    Guimarães-Costa, Anderson B.; Nascimento, Michelle T. C.; Froment, Giselle S.; Soares, Rodrigo P. P.; Morgado, Fernanda N.; Conceição-Silva, Fátima; Saraiva, Elvira M.

    2009-01-01

    Neutrophils are short-lived leukocytes that die by apoptosis, necrosis, and NETosis. Upon death by NETosis, neutrophils release fibrous traps of DNA, histones, and granule proteins named neutrophil extracellular traps (NETs), which can kill bacteria and fungi. Inoculation of the protozoan Leishmania into the mammalian skin causes local inflammation with neutrophil recruitment. Here, we investigated the release of NETs by human neutrophils upon their interaction with Leishmania parasites and NETs' ability to kill this protozoan. The NET constituents DNA, elastase, and histones were detected in traps associated to promastigotes by immunofluorescence. Electron microscopy revealed that Leishmania was ensnared by NETs released by neutrophils. Moreover, Leishmania and its surface lipophosphoglycan induced NET release by neutrophils in a parasite number- and dose-dependent manner. Disruption of NETs by DNase treatment during Leishmania–neutrophil interaction increased parasite survival, evidencing NETs' leishmanicidal effect. Leishmania killing was also elicited by NET-rich supernatants from phorbol 12-myristate 13-acetate-activated neutrophils. Immunoneutralization of histone during Leishmania–neutrophil interaction partially reverted Leishmania killing, and purified histone killed the parasites. Meshes composed of DNA and elastase were evidenced in biopsies of human cutaneous leishmaniasis. NET is an innate response that might contribute to diminish parasite burden in the Leishmania inoculation site. PMID:19346483

  16. Choline transport in Leishmania major promastigotes and its inhibition by choline and phosphocholine analogs.

    PubMed

    Zufferey, Rachel; Mamoun, Choukri Ben

    2002-01-01

    Phosphatidylcholine is the most abundant phospholipid in the membranes of the human parasite Leishmania. The metabolic pathways leading to its biosynthesis are likely to play a critical role in parasite development and survival and may offer a good target for antileishmanial chemotherapy. Phosphatidylcholine synthesis via the CDP-choline pathway requires transport of the choline precursor from the host. Here, we report the first characterization of choline transport in this parasite, which is carrier-mediated and exhibits Michaelis-Menten kinetics with an apparent K(m) value of 2.5 microM for choline. This process is Na(+)-independent and requires an intact proton gradient to be fully functional. Choline transport into Leishmania is highly specific for choline and is inhibited by the choline carrier inhibitor hemicholinium-3, the channel blocker quinacrine, the antimalarial aminoquinolines quinine and quinidine, the antileishmanial phosphocholine analogs, miltefosine and edelfosine, and by choline analogs, most of which have antimalarial activities. Most importantly, choline analogs kill the promastigote form of the parasite in vitro in the low micromolar range. These results set the stage for the use of choline analogs in antileishmanial chemotherapy and shed new lights on the mechanism of action of the leishmanicidal phosphocholine analogs. PMID:12467980

  17. The Vanishing Mexicana/o: (Dis)Locating the Native in Ruiz de Burton's "Who Would Have thought It?" and "The Squatter and the Don"

    ERIC Educational Resources Information Center

    Szeghi, Tereza M.

    2011-01-01

    This article complements the existing body of Ruiz de Burton scholarship by providing the first sustained examination of her literary representations of American Indians in both "Who Would Have Thought It?" (1872) and "The Squatter and the Don" (1885), and by exploring how these representations serve her broader aims of social and political…

  18. Migrant Mexican Traditions = Tradiciones Migrantes Mexicanas. An Exhibit of Folk Art by Mexican Migrant Farmworkers (Geneseo, New York, September 22-October 4, 1990).

    ERIC Educational Resources Information Center

    Camacho, Juana; Gomez, Jose Luis

    This exhibit guide (in Spanish, with translation in English printed on adjoining columns on each page), describes an exhibition of folk art by Mexican migrant farmworkers presented by thre Folk Arts Program of the BOCES Geneseo Migrant Center. The exhibit is divided into four major themes that farmworkers presented by the BOCES Geneseo Migrant…

  19. RNA editing and mitochondrial activity in promastigotes and amastigotes of Leishmania donovani✯

    PubMed Central

    Neboháčová, Martina; Kim, Christine E.; Simpson, Larry; Maslov, Dmitri A.

    2009-01-01

    Kinetoplast maxicircle DNA sequence organization was investigated in Leishmania donovani, strain 1S LdBob. Gene arrangement in the coding (conserved) region of the maxicircle is collinear with that of most trypanosomatids, with individual genes showing 80-90% nucleotide identity to Leishmania tarentolae, strain UC. The notable exception was an integration of a full-size minicircle sequence in the ND1 gene coding region found in L. donovani. Editing patterns of the mitochondrial mRNAs investigated also followed L. tarentolae UC patterns, including productive editing of the components of respiratory complexes III, IV and V, and ribosomal protein S12 (RPS12), as well as the lack of productive editing in five out of six pan-edited cryptogenes (ND3, ND8, ND9, G3, G4) found in these species. Several guide RNAs for the editing events were localized in minicircles and maxicircles in the locations that are conserved between the species. Mitochondrial activity, including rates of oxygen consumption, the presence and the levels of respiratory complexes and their individual subunits and the steady-state levels of several mitochondrial-encoded mRNAs were essentially the same in axenically grown amastigotes and in promastigotes of L. donovani. However, some modulation of mitochondrial activity between these developmental stages was suggested by the finding of an amastigote-specific component in Complex IV, a down-regulation of mitochondrial RNA-binding proteins (MRP) [define] and MRP-associated protein (MRP-AP) in amastigotes, and by variations in the levels of RPS12, ND3, ND9, G3 and G4 pre-edited transcripts. PMID:19109964

  20. Ca2+ transport in isolated mitochondrial vesicles from Leishmania braziliensis promastigotes.

    PubMed

    Benaim, G; Bermudez, R; Urbina, J A

    1990-02-01

    Leishmania braziliensis maintained very low (50 +/- 20 nM) intracellular concentrations of calcium ions under normal conditions, as shown by the fluorimetric indicator QUIN2. Digitonin-permeabilized cells liberated large amounts of calcium ions in the presence of the ionophore A23187, indicating the presence of a large intracellular reservoir for this ion. Given the extraordinary extension of the single giant mitochondrion of Kinetoplastida and the known capacity of mitochondria from other sources to accumulate calcium, we tested the capacity of this organelle to accumulate calcium ions in Leishmania. Coupled mitochondrial vesicles, five-fold enriched in succinate-cytochrome c oxidoreductase, were obtained from promastigotes by gentle grinding (45 s) with glass beads in hypertonic buffer solution, followed by differential centrifugation. These vesicles had a respiratory control ratio of 1.82 +/- 0.15, and two phosphorylation sites (sites II and III) using succinate as electron donor, and were capable of calcium uptake in the presence of several respiratory substrates; this uptake was enhanced in the presence of ADP and Pi and was blocked by classical electron transport inhibitors. Uncouplers such as carbonyl cyanide p-trifluoromethoxy-phenylhydrazone (FCCP) and the calcium ionophore A23187 released previously accumulated calcium ions, suggesting that the driving force for the calcium uptake by the vesicles is the respiratory generated electrochemical potential gradient of protons. A study of the affinity of this system for calcium showed that even at 90 microM free calcium, succinate-induced calcium uptake is not saturated while approaching a level of 200 nmol min-1 (mg protein)-1, indicating a low-affinity, large-capacity system.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2304488

  1. Recombinant Forms of Leishmania amazonensis Excreted/Secreted Promastigote Surface Antigen (PSA) Induce Protective Immune Responses in Dogs.

    PubMed

    Petitdidier, Elodie; Pagniez, Julie; Papierok, Gérard; Vincendeau, Philippe; Lemesre, Jean-Loup; Bras-Gonçalves, Rachel

    2016-05-01

    Preventive vaccination is a highly promising strategy for interrupting leishmaniasis transmission that can, additionally, contribute to elimination. A vaccine formulation based on naturally excreted secreted (ES) antigens was prepared from L. infantum promastigote culture supernatant. This vaccine achieved successful results in Phase III trials and was licensed and marketed as CaniLeish. We recently showed that newly identified ES promastigote surface antigen (PSA), from both viable promastigotes and axenically-grown amastigotes, represented the major constituent and the highly immunogenic antigen of L. infantum and L. amazonensis ES products. We report here that three immunizations with either the recombinant ES LaPSA-38S (rPSA) or its carboxy terminal part LaPSA-12S (Cter-rPSA), combined with QA-21 as adjuvant, confer high levels of protection in naive L. infantum-infected Beagle dogs, as checked by bone marrow parasite absence in respectively 78.8% and 80% of vaccinated dogs at 6 months post-challenge. The parasite burden in infected vaccinated dogs was significantly reduced compared to placebo group, as measured by q-PCR. Moreover, our results reveal humoral and cellular immune response clear-cut differences between vaccinated and control dogs. An early increase in specific IgG2 antibodies was observed in rPSA/QA-21- and Cter-rPSA/QA-21-immunized dogs only. They were found functionally active in vitro and were highly correlated with vaccine protection. In vaccinated protected dogs, IFN-γ and NO productions, as well as anti-leishmanial macrophage activity, were increased. These data strongly suggest that ES PSA or its carboxy-terminal part, in recombinant forms, induce protection in a canine model of zoonotic visceral leishmaniasis by inducing a Th1-dominant immune response and an appropriate specific antibody response. These data suggest that they could be considered as important active components in vaccine candidates. PMID:27223609

  2. Recombinant Forms of Leishmania amazonensis Excreted/Secreted Promastigote Surface Antigen (PSA) Induce Protective Immune Responses in Dogs

    PubMed Central

    Petitdidier, Elodie; Pagniez, Julie; Papierok, Gérard; Vincendeau, Philippe; Lemesre, Jean-Loup; Bras-Gonçalves, Rachel

    2016-01-01

    Preventive vaccination is a highly promising strategy for interrupting leishmaniasis transmission that can, additionally, contribute to elimination. A vaccine formulation based on naturally excreted secreted (ES) antigens was prepared from L. infantum promastigote culture supernatant. This vaccine achieved successful results in Phase III trials and was licensed and marketed as CaniLeish. We recently showed that newly identified ES promastigote surface antigen (PSA), from both viable promastigotes and axenically-grown amastigotes, represented the major constituent and the highly immunogenic antigen of L. infantum and L. amazonensis ES products. We report here that three immunizations with either the recombinant ES LaPSA-38S (rPSA) or its carboxy terminal part LaPSA-12S (Cter-rPSA), combined with QA-21 as adjuvant, confer high levels of protection in naive L. infantum-infected Beagle dogs, as checked by bone marrow parasite absence in respectively 78.8% and 80% of vaccinated dogs at 6 months post-challenge. The parasite burden in infected vaccinated dogs was significantly reduced compared to placebo group, as measured by q-PCR. Moreover, our results reveal humoral and cellular immune response clear-cut differences between vaccinated and control dogs. An early increase in specific IgG2 antibodies was observed in rPSA/QA-21- and Cter-rPSA/QA-21-immunized dogs only. They were found functionally active in vitro and were highly correlated with vaccine protection. In vaccinated protected dogs, IFN-γ and NO productions, as well as anti-leishmanial macrophage activity, were increased. These data strongly suggest that ES PSA or its carboxy-terminal part, in recombinant forms, induce protection in a canine model of zoonotic visceral leishmaniasis by inducing a Th1-dominant immune response and an appropriate specific antibody response. These data suggest that they could be considered as important active components in vaccine candidates. PMID:27223609

  3. Cellular Growth and Mitochondrial Ultrastructure of Leishmania (Viannia) braziliensis Promastigotes Are Affected by the Iron Chelator 2,2-Dipyridyl

    PubMed Central

    Mesquita-Rodrigues, Camila; Menna-Barreto, Rubem F. S.; Sabóia-Vahia, Leonardo; Da-Silva, Silvia A. G.; de Souza, Elen M.; Waghabi, Mariana C.; Cuervo, Patrícia; De Jesus, José B.

    2013-01-01

    Background Iron is an essential element for the survival of microorganisms in vitro and in vivo, acting as a cofactor of several enzymes and playing a critical role in host-parasite relationships. Leishmania (Viannia) braziliensis is a parasite that is widespread in the new world and considered the major etiological agent of American tegumentary leishmaniasis. Although iron depletion leads to promastigote and amastigote growth inhibition, little is known about the role of iron in the biology of Leishmania. Furthermore, there are no reports regarding the importance of iron for L. (V.) braziliensis. Methodology/Principal Findings In this study, the effect of iron on the growth, ultrastructure and protein expression of L. (V.) braziliensis was analyzed by the use of the chelator 2,2-dipyridyl. Treatment with 2,2-dipyridyl affected parasites' growth in a dose- and time-dependent manner. Multiplication of the parasites was recovered after reinoculation in fresh culture medium. Ultrastructural analysis of treated promastigotes revealed marked mitochondrial swelling with loss of cristae and matrix and the presence of concentric membranar structures inside the organelle. Iron depletion also induced Golgi disruption and intense cytoplasmic vacuolization. Fluorescence-activated cell sorting analysis of tetramethylrhodamine ester-stained parasites showed that 2,2-dipyridyl collapsed the mitochondrial membrane potential. The incubation of parasites with propidium iodide demonstrated that disruption of mitochondrial membrane potential was not associated with plasma membrane permeabilization. TUNEL assays indicated no DNA fragmentation in chelator-treated promastigotes. In addition, two-dimensional electrophoresis showed that treatment with the iron chelator induced up- or down-regulation of proteins involved in metabolism of nucleic acids and coordination of post-translational modifications, without altering their mRNA levels. Conclusions Iron chelation leads to a

  4. Immunomodulatory Impact of Leishmania-Induced Macrophage Exosomes: A Comparative Proteomic and Functional Analysis

    PubMed Central

    Hassani, Kasra; Olivier, Martin

    2013-01-01

    Released by many eukaryotic cells, the exosomes are 40–100 nm vesicles shown to operate over the complex processes of cell-cell communication. Among the metazoan cell lineages known to generate exosomes is the mononuclear phagocyte lineage, a lineage that parasites such as Leishmania are known to subvert as host cells. We previously reported that mouse macrophage signaling and functions are modified once co-incubated with exoproteome of Leishmania promastigotes. Using mass spectrometry analysis, we were curious to further compare the content of purified exosomes released by the J774 mouse macrophage cell line exposed or not to either LPS or to stationary phase Leishmania mexicana promastigotes. Collectively, our analyses resulted in detection of 248 proteins, ∼50–80% of which were shared among the three sources studied. Using exponentially modified protein abundance index (emPAI) and network analyses, we found that the macrophage exosomes display unique signatures with respect to composition and abundance of many functional groups of proteins, such as plasma membrane-associated proteins, chaperones and metabolic enzymes. Moreover, for the first time, L. mexicana surface protease GP63 is shown to be present in exosomes released from J774 macrophages exposed to stationary phase promastigotes. We observed that macrophage exosomes are able to induce signaling molecules and transcription factors in naive macrophages. Finally, using qRT-PCR, we monitored modulation of expression of multiple immune-related genes within macrophages exposed to exosomes. We found all three groups of exosomes to induce expression of immune-related genes, the ones collected from macrophages exposed to L. mexicana sharing properties with exosomes collected from macrophage left unexposed to any agonist. Overall, our results allowed depicting that protein sorting into macrophage-derived exosomes depends upon the cell status and how such distinct protein sorting can in turn impact the

  5. Combined effect of the essential oil from Chenopodium ambrosioides and antileishmanial drugs on promastigotes of Leishmania amazonensis.

    PubMed

    Monzote, Lianet; Montalvo, Ana Margarita; Scull, Ramón; Miranda, Migdalia; Abreu, Juan

    2007-01-01

    To date, there are no vaccines against Leishmania, and chemotherapy remains the mainstay for the control of leishmaniasis. The drugs of choice used for leishmaniasis therapy are significantly toxic, expensive and with a growing frequency of refractory infections. Because of these limitations, a combination therapy is the better hope. This work demonstrates that the essential oil from Chenopodium ambrosioides shows a synergic activity after incubation in conjunction with pentamidine against promastigotes of Leishmania amazonensis. However, an indifferent effect has been found for combinations of meglumine antimoniate or amphotericin B and the essential oil. PMID:17823757

  6. Antileishmanial activity of quinazoline derivatives: synthesis, docking screens, molecular dynamic simulations and electrochemical studies.

    PubMed

    Mendoza-Martínez, Cesar; Galindo-Sevilla, Norma; Correa-Basurto, José; Ugalde-Saldivar, Victor Manuel; Rodríguez-Delgado, Rosa Georgina; Hernández-Pineda, Jessica; Padierna-Mota, Cecilia; Flores-Alamo, Marcos; Hernández-Luis, Francisco

    2015-03-01

    A series of quinazoline-2,4,6-triamine were synthesized and evaluated in vitro against Leishmania mexicana. Among them, N(6)-(ferrocenmethyl)quinazolin-2,4,6-triamine (H2) showed activity on promastigotes and intracellular amastigotes, as well as low cytotoxicity in mammalian cells. Docking and electrochemical studies showed the importance of both the ferrocene and the heterocyclic nucleus to the observed activity. H2 is readily oxidized electrochemically, indicating that the mechanism of action probably involves redox reactions. PMID:25576738

  7. Deprivation of L-Arginine Induces Oxidative Stress Mediated Apoptosis in Leishmania donovani Promastigotes: Contribution of the Polyamine Pathway

    PubMed Central

    Mandal, Abhishek; Das, Sushmita; Roy, Saptarshi; Ghosh, Ayan Kumar; Sardar, Abul Hasan; Verma, Sudha; Saini, Savita; Singh, Ruby; Abhishek, Kumar; Kumar, Ajay; Mandal, Chitra; Das, Pradeep

    2016-01-01

    The growth and survival of intracellular parasites depends on the availability of extracellular nutrients. Deprivation of nutrients viz glucose or amino acid alters redox balance in mammalian cells as well as some lower organisms. To further understand the relationship, the mechanistic role of L-arginine in regulation of redox mediated survival of Leishmania donovani promastigotes was investigated. L-arginine deprivation from the culture medium was found to inhibit cell growth, reduce proliferation and increase L-arginine uptake. Relative expression of enzymes, involved in L-arginine metabolism, which leads to polyamine and trypanothione biosynthesis, were downregulated causing decreased production of polyamines in L-arginine deprived parasites and cell death. The resultant increase in reactive oxygen species (ROS), due to L-arginine deprivation, correlated with increased NADP+/NADPH ratio, decreased superoxide dismutase (SOD) level, increased lipid peroxidation and reduced thiol content. A deficiency of L-arginine triggered phosphatidyl serine externalization, a change in mitochondrial membrane potential, release of intracellular calcium and cytochrome-c. This finally led to DNA damage in Leishmania promastigotes. In summary, the growth and survival of Leishmania depends on the availability of extracellular L-arginine. In its absence the parasite undergoes ROS mediated, caspase-independent apoptosis-like cell death. Therefore, L-arginine metabolism pathway could be a probable target for controlling the growth of Leishmania parasites and disease pathogenesis. PMID:26808657

  8. Identification of cross-reactive promastigote cell surface antigens of some leishmanial stocks by 125I labeling and immunoprecipitation.

    PubMed Central

    Gardiner, P R; Jaffe, C L; Dwyer, D M

    1984-01-01

    Externally oriented surface membrane constituents of promastigotes from several Leishmania species were radiolabeled with 125I. Autoradiographs of cell surface-labeled and sodium dodecyl sulfate-polyacrylamide gel electrophoresis-separated proteins of the stocks revealed distinctive patterns of bands in the molecular weight range of 6,000 to 240,000. Immunoprecipitation of detergent extracts of the labeled promastigote stocks with anti-Leishmania donovani membrane serum demonstrated that each of the stocks contained some antigenically cross-reactive determinants. The electrophoretic patterns of these determinants serve both to distinguish the parasite stocks (by unique, species-specific patterns) and to indicate antigenic similarities in stocks thought to be different by other biochemical criteria. At least 12 cross-reactive cell surface antigens in two New World leishmanias are recognized by polyvalent anti-L. donovani serum, suggesting that these common leishmanial antigens may account for the documented serological cross-reactivities among various Leishmania species. In all stocks tested, an iodinated protein was identified which had a relative molecular weight of 65,000 under reducing conditions but which demonstrated an increase in relative mobility in sodium dodecyl sulfate-polyacrylamide gels under nonreducing conditions. Distinctive patterns of the antigens common to the several stocks were also demonstrated with the use of monoclonal antibodies. Images PMID:6363295

  9. Deprivation of L-Arginine Induces Oxidative Stress Mediated Apoptosis in Leishmania donovani Promastigotes: Contribution of the Polyamine Pathway.

    PubMed

    Mandal, Abhishek; Das, Sushmita; Roy, Saptarshi; Ghosh, Ayan Kumar; Sardar, Abul Hasan; Verma, Sudha; Saini, Savita; Singh, Ruby; Abhishek, Kumar; Kumar, Ajay; Mandal, Chitra; Das, Pradeep

    2016-01-01

    The growth and survival of intracellular parasites depends on the availability of extracellular nutrients. Deprivation of nutrients viz glucose or amino acid alters redox balance in mammalian cells as well as some lower organisms. To further understand the relationship, the mechanistic role of L-arginine in regulation of redox mediated survival of Leishmania donovani promastigotes was investigated. L-arginine deprivation from the culture medium was found to inhibit cell growth, reduce proliferation and increase L-arginine uptake. Relative expression of enzymes, involved in L-arginine metabolism, which leads to polyamine and trypanothione biosynthesis, were downregulated causing decreased production of polyamines in L-arginine deprived parasites and cell death. The resultant increase in reactive oxygen species (ROS), due to L-arginine deprivation, correlated with increased NADP+/NADPH ratio, decreased superoxide dismutase (SOD) level, increased lipid peroxidation and reduced thiol content. A deficiency of L-arginine triggered phosphatidyl serine externalization, a change in mitochondrial membrane potential, release of intracellular calcium and cytochrome-c. This finally led to DNA damage in Leishmania promastigotes. In summary, the growth and survival of Leishmania depends on the availability of extracellular L-arginine. In its absence the parasite undergoes ROS mediated, caspase-independent apoptosis-like cell death. Therefore, L-arginine metabolism pathway could be a probable target for controlling the growth of Leishmania parasites and disease pathogenesis. PMID:26808657

  10. Identification of Leishmania spp. promastigotes in the intestines, ovaries, and salivary glands of Rhipicephalus sanguineus actively infesting dogs.

    PubMed

    Viol, Milena Araúz; Guerrero, Felix D; de Oliveira, Bruno César Miranda; de Aquino, Monally Conceição Costa; Loiola, Saulo Hudson; de Melo, Guilherme Dias; de Souza Gomes, Aparecida Helena; Kanamura, Cristina Takami; Garcia, Marcos Valério; Andreotti, Renato; de Lima, Valéria Marçal Félix; Bresciani, Katia Denise Saraiva

    2016-09-01

    Sand flies are recognized as the major vector of canine visceral leishmaniasis. However, in some areas of Brazil where sand flies do not occur, this disease is found in humans and dogs. There has been speculation that ticks might play a role in transmission of canine visceral leishmaniasis and the DNA of Leishmania spp. has been reported in whole ticks. We investigated the presence of Leishmania spp. promastigotes in the intestines, ovaries, and salivary glands of Rhipicephalus sanguineus ticks collected from tick-infested dogs in two cities of Brazil. We used 66 dogs that tested positive and 33 that tested negative for Leishmania spp. according to direct cytological examination assays. Ten ticks were collected from each dog and dissected to collect the intestines, ovaries, and salivary glands for immunohistochemistry (IHC) and diagnostic real-time polymerase chain reaction (RT-PCR). IHC results showed Leishmania spp. in 98, 14, and 8 % of the intestines, ovaries, and salivary glands, respectively. Real-time PCR showed that 89, 41, and 33 % of the tick intestine, ovary, and salivary glands, respectively, were positive for Leishmania spp. The verification of promastigotes of Leishmania spp. by two independent techniques in ticks collected from these urban region dogs showed that there is need for clarification of the role of ticks in the transmission of canine visceral leishmaniasis in Brazil. PMID:27169724

  11. In vitro selection of miltefosine resistance in promastigotes of Leishmania donovani from Nepal: genomic and metabolomic characterization

    PubMed Central

    Shaw, C. D.; Lonchamp, J.; Downing, T.; Imamura, H.; Freeman, T. M.; Cotton, J. A.; Sanders, M.; Blackburn, G.; Dujardin, J. C.; Rijal, S.; Khanal, B.; Illingworth, C. J. R.; Coombs, G. H.

    2016-01-01

    Summary In this study, we followed the genomic, lipidomic and metabolomic changes associated with the selection of miltefosine (MIL) resistance in two clinically derived Leishmania donovani strains with different inherent resistance to antimonial drugs (antimony sensitive strain Sb‐S; and antimony resistant Sb‐R). MIL‐R was easily induced in both strains using the promastigote‐stage, but a significant increase in MIL‐R in the intracellular amastigote compared to the corresponding wild‐type did not occur until promastigotes had adapted to 12.2 μM MIL. A variety of common and strain‐specific genetic changes were discovered in MIL‐adapted parasites, including deletions at the LdMT transporter gene, single‐base mutations and changes in somy. The most obvious lipid changes in MIL‐R promastigotes occurred to phosphatidylcholines and lysophosphatidylcholines and results indicate that the Kennedy pathway is involved in MIL resistance. The inherent Sb resistance of the parasite had an impact on the changes that occurred in MIL‐R parasites, with more genetic changes occurring in Sb‐R compared with Sb‐S parasites. Initial interpretation of the changes identified in this study does not support synergies with Sb‐R in the mechanisms of MIL resistance, though this requires an enhanced understanding of the parasite's biochemical pathways and how they are genetically regulated to be verified fully. PMID:26713880

  12. Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): localization and in vitro inhibition of promastigotes growth by polyclonal antibodies.

    PubMed

    Porcino, Gabriane Nascimento; Carvalho-Campos, Cristiane; Maia, Ana Carolina Ribeiro Gomes; Detoni, Michelle Lima; Faria-Pinto, Priscila; Coimbra, Elaine Soares; Marques, Marcos José; Juliano, Maria Aparecida; Juliano, Luiz; Diniz, Vanessa Álvaro; Corte-Real, Suzana; Vasconcelos, Eveline Gomes

    2012-10-01

    Nucleoside triphosphate diphosphohydrolase (NTPDase) activity was recently characterized in Leishmania (Viannia) braziliensis promastigotes (Lb), and an antigenic conserved domain (r82-121) from the specific NTPDase 1 isoform was identified. In this work, mouse polyclonal antibodies produced against two synthetic peptides derived from this domain (LbB1LJ, r82-103; LbB2LJ, r102-121) were used. The anti-LbB1LJ or anti-LbB2LJ antibodies were immobilized on protein A-sepharose and immunoprecipitated the NTPDase 1 of 48 kDa and depleted approximately 40% of the phosphohydrolytic activity from detergent-homogenized Lb preparation. Ultrastructural immunocytochemical microscopy identified the NTPDase 1 on the parasite surface and in its subcellular cytoplasmic vesicles, mitochondria, kinetoplast and nucleus. The ATPase and ADPase activities of detergent-homogenized Lb preparation were partially inhibited by anti-LbB1LJ antibody (43-79%), which was more effective than that inhibition (18-47%) by anti-LbB2LJ antibody. In addition, the immune serum anti-LbB1LJ (67%) or anti-LbB2LJ (33%) was cytotoxic, significantly reducing the promastigotes growth in vitro. The results appoint the conserved domain from the L. braziliensis NTPDase as an important target for inhibitor design and the potential application of these biomolecules in experimental protocols of disease control. PMID:22921497

  13. Metabolomics-Based Study of Logarithmic and Stationary Phases of Promastigotes in Leishmania major by 1H NMR Spectroscopy

    PubMed Central

    Arjmand, Mohammad; Madrakian, Azadeh; Khalili, Ghader; Najafi, Ali; Zamani, Zahra; Akbari, Ziba

    2016-01-01

    Background: Cutaneous leishmaniasis is one of the most important parasitic diseases in humans. In this disease, one of the responsible organisms is Leishmania major, which is transmitted by sandfly vector. There are specific differences in biochemical profiles and metabolite pathways in logarithmic and stationary phases of Leishmania parasites. In the present study, 1H NMR spectroscopy was used to examine the metabolites outliers in the logarithmic and stationary phases of promastigotes in L. major to enlighten more about the transmission mechanism in metacyclogenesis of L. major. Methods: Promastigote was cultured, logarithmic and stationary phases were separated by the peanut agglutinin, and cell metabolites were extracted. 1H NMR spectroscopy was applied, and outliers were analyzed using principal component analysis. Results: The most altered metabolites in stationary and logarithmic phases were limited to citraconic acid, isopropylmalic acid, L-leucine, ornithine, caprylic acid, capric acid, and acetic acid. Conclusion: 1H NMR spectroscopy could play an important role in the characterization of metabolites in biochemical pathways during a metacyclogenesis process. These metabolites and their pathways can help in exploiting a transmission mechanism in metacyclogenesis, and outcoming data might be used in the metabolic network reconstruction of L. major modeling. PMID:26592771

  14. Adoptive transfer of dendritic cells modulates immunogenesis and tolerogenesis in a neonatal model of murine cutaneous leishmaniasis

    PubMed Central

    Ponce, Loida V; Corado, José; Díaz, Nilka L; Tapia, Felix J

    2005-01-01

    We evaluated the adoptive transfer of DCs on Leishmania (L.) mexicana-infected neonatal BALB/c mice. DCs were isolated and purified from the spleens of the following donor groups: a) Adult BALB/c mice infected during adulthood with L. (L) mexicana; b) Adult BALB/c mice infected during neonatal life; c) Healthy neonatal BALB/c mice; d) Healthy adult BALB/c mice. A neonatal model of infection, generated after inoculation with 5 × 105 promastigotes of L. (L) mexicana, was used as the infection control group. Sixteen hours after intraperitoneal transfer of DCs (1 × 103, 1 × 105, or 1 × 106 cells/ml), neonatal recipient BALB/c mice were infected. The adoptive transfer of DCs diminished disease progression in neonatal mice. This reduction depends on the quantity and provenance of transferred DCs, since the effect was more evident with high numbers of DCs from adult mice infected during adulthood and healthy neonatal mice. Protection was significantly reduced in animals receiving DCs from healthy adult mice but it was absent in mice receiving DCs from adult mice infected during neonatal life. These results suggest that genetic susceptibility to Leishmania infection can be modified during neonatal life, and that the period of life when antigens are encountered is crucial in influencing the capacity of DCs to induce resistance or tolerance. PMID:15670331

  15. Antileishmanial activity of a mixture of Tridax procumbens and Allium sativum in mice

    PubMed Central

    Gamboa-Leon, Rubi; Vera-Ku, Marina; Peraza-Sanchez, Sergio R.; Ku-Chulim, Carlos; Horta-Baas, Aurelio; Rosado-Vallado, Miguel

    2014-01-01

    We tested a mixture of Tridax procumbens, known for its direct action against Leishmania mexicana, and Allium sativum, known for its immunomodulatory effect, as an alternative to treat cutaneous leishmaniasis. Acute oral toxicity was tested with the Up-and-Down Procedure (UDP) using a group of healthy mice administered with either T. procumbens or A. sativum extracts and compared with a control group. Liver injury and other parameters of toxicity were determined in mice at day 14. The in vivo assay was performed with mice infected with L. mexicana promastigotes and treated with either a mixture of T. procumbens and A. sativum or each extract separately. The thickness of the mice’s footpads was measured weekly. After the 12-week period of infection, blood samples were obtained by cardiac puncture to determine the total IgG, IgG1 and IgG2a immunoglobulins by a noncommercial indirect ELISA. We showed that the mixture of T. procumbens and A. sativum extracts was better at controlling L. mexicana infection while not being toxic when tested in the acute oral toxicity assay in mice. An increase in the ratio of IgG2a/IgG1 indicated a tendency to raise a Th1-type immune response in mice treated with the mixture. The mixture of T. procumbens and A. sativum extracts is a promising natural treatment for cutaneous leishmaniasis and its healing effects make it a good candidate for a possible new phytomedicine. PMID:24717526

  16. Proteophosophoglycans Regurgitated by Leishmania-Infected Sand Flies Target the L-Arginine Metabolism of Host Macrophages to Promote Parasite Survival

    PubMed Central

    Rogers, Matthew; Kropf, Pascale; Choi, Beak-San; Dillon, Rod; Podinovskaia, Maria; Bates, Paul; Müller, Ingrid

    2009-01-01

    All natural Leishmania infections start in the skin; however, little is known of the contribution made by the sand fly vector to the earliest events in mammalian infection, especially in inflamed skin that can rapidly kill invading parasites. During transmission sand flies regurgitate a proteophosphoglycan gel synthesized by the parasites inside the fly midgut, termed promastigote secretory gel (PSG). Regurgitated PSG can exacerbate cutaneous leishmaniasis. Here, we show that the amount of Leishmania mexicana PSG regurgitated by Lutzomyia longipalpis sand flies is proportional to the size of its original midgut infection and the number of parasites transmitted. Furthermore, PSG could exacerbate cutaneous L. mexicana infection for a wide range of doses (10–10,000 parasites) and enhance infection by as early as 48 hours in inflamed dermal air pouches. This early exacerbation was attributed to two fundamental properties of PSG: Firstly, PSG powerfully recruited macrophages to the dermal site of infection within 24 hours. Secondly, PSG enhanced alternative activation and arginase activity of host macrophages, thereby increasing L-arginine catabolism and the synthesis of polyamines essential for intracellular parasite growth. The increase in arginase activity promoted the intracellular growth of L. mexicana within classically activated macrophages, and inhibition of macrophage arginase completely ablated the early exacerbatory properties of PSG in vitro and in vivo. Thus, PSG is an essential component of the infectious sand fly bite for the early establishment of Leishmania in skin, which should be considered when designing and screening therapies against leishmaniasis. PMID:19696894

  17. The proliferation potential of promastigotes of the main Leishmania species of the old world in NNN culture medium prepared using blood of four different mammals.

    PubMed

    Ladopoulos, Theodoros; Ntais, Pantelis; Tsirigotakis, Nikolaos; Dokianakis, Emmanouil; Antoniou, Maria

    2015-10-01

    The efficacy of the in vitro cultivation of promastigotes of four Leishmania spp. was tested in the biphasic Novy-MacNeal-Nicolle (NNN) medium prepared using blood from different animals (horse, donkey, goat and sheep). The aim was to test which NNN preparation gave the best yield in the shortest time for different parasite species, in order to obtain a large crop of promastigotes for experimental work and for antigen preparation. Promastigotes of Leishmania infantum, Leishmania donovani, Leishmania tropica and Leishmania major, the four main parasite species occurring in the old world, were defrosted from -80 °C and placed, at equal numbers, in the 4 different NNN preparations. At the end of the 7th day, the NNN medium using horse blood produced the greatest number of promastigotes for all Leishmania spp. tested, whilst goat blood proved the poorest medium, providing culture results only for L. infantum. This finding may be explained by the fact that Leishmania is a nicotinamide adenine dinucleotide (NAD) auxotroph and horse erythrocytes support NAD-dependent microorganisms. PMID:26219203

  18. Ecto-Nucleotidase Activities of Promastigotes from Leishmania (Viannia) braziliensis Relates to Parasite Infectivity and Disease Clinical Outcome

    PubMed Central

    Leite, Pauline M.; Gomes, Rodrigo S.; Figueiredo, Amanda B.; Serafim, Tiago D.; Tafuri, Wagner L.; de Souza, Carolina C.; Moura, Sandra A. L.; Fietto, Juliana L. R.; Melo, Maria N.; Ribeiro-Dias, Fátima; Oliveira, Milton A. P.; Rabello, Ana; Afonso, Luís C. C.

    2012-01-01

    Background Leishmania (Viannia) braziliensis has been associated with a broad range of clinical manifestations ranging from a simple cutaneous ulcer to destructive mucosal lesions. Factors leading to this diversity of clinical presentations are not clear, but parasite factors have lately been recognized as important in determining disease progression. Given the fact that the activity of ecto-nucleotidases correlates with parasitism and the development of infection, we evaluated the activity of these enzymes in promastigotes from 23 L. braziliensis isolates as a possible parasite-related factor that could influence the clinical outcome of the disease. Methodology/Principal Findings Our results show that the isolates differ in their ability to hydrolyze adenine nucleotides. Furthermore, we observed a positive correlation between the time for peak of lesion development in C57BL/6J mice and enzymatic activity and clinical manifestation of the isolate. In addition, we found that L. (V.) braziliensis isolates obtained from mucosal lesions hydrolyze higher amounts of adenine nucleotides than isolates obtained from skin lesions. One isolate with high (PPS6m) and another with low (SSF) ecto-nucleotidase activity were chosen for further studies. Mice inoculated with PPS6m show delayed lesion development and present larger parasite loads than animals inoculated with the SSF isolate. In addition, PPS6m modulates the host immune response by inhibiting dendritic cell activation and NO production by activated J774 macrophages. Finally, we observed that the amastigote forms from PPS6m and SSF isolates present low enzymatic activity that does not interfere with NO production and parasite survival in macrophages. Conclusions/Significance Our data suggest that ecto-nucleotidases present on the promastigote forms of the parasite may interfere with the establishment of the immune response with consequent impaired ability to control parasite dissemination and this may be an important

  19. Identification of developmentally-regulated proteins in Leishmania panamensis by proteome profiling of promastigotes and axenic amastigotes.

    PubMed

    Walker, John; Vasquez, Juan-José; Gomez, Maria Adelaida; Drummelsmith, Jolyne; Burchmore, Richard; Girard, Isabelle; Ouellette, Marc

    2006-05-01

    We have employed proteomics to identify proteins upregulated in the amastigote life-stage of Leishmaniapanamensis, using axenically-differentiated forms as models of authentic intracellular parasites. Resolution of the soluble proteomes of axenic amastigotes and promastigotes by two-dimensional electrophoresis (2DE) in the neutral pI range (5-7) revealed equivalent numbers of protein spots in both life-stages (644-682 using Coomassie Blue and 851-863 by silver staining). Although representing a relatively low proportion (8.1-10.8%) of the predicted 8000 gene products of Leishmania, these proteome maps enabled the reproducible detection of 75 differentially-regulated protein spots in amastigotes, comprising 24 spots "uniquely" expressed in this life-stage and 51 over-expressed by 1.2-5.7-fold compared to promastigotes. Of the 11 amastigote-specific spots analysed by mass spectrometry (MS), 5 yielded peptide sequences with no orthologues in Leishmania major, and the remaining 6 were identified as 7 distinct proteins (some of which were truncated isoforms) representing several functional classes: carbohydrate/energy metabolism (fructose 1,6-bisphosphate aldolase, glucose 6-phosphate dehydrogenase, pyruvate dehydrogenase), stress response (heat shock protein [HSP] 83), cell membrane/cytoskeleton (beta-tubulin), amino acid metabolism (cysteine synthase) and cell-cycle (ran-binding protein). Four additional over-expressed spots were tentatively identified as HSPs 60 and 70 and HSP 70-related proteins -1 and -4 by positional analogy with these landmark proteins in the Leishmania guyanensis proteome. Our data demonstrate the feasibility of proteomics as an approach to identify novel developmentally-regulated proteins linked to Leishmania differentiation and intracellular survival, while simultaneously pinpointing therapeutic targets. In particular, the amastigote-specific expression of cysteine synthase underlines the importance of de novo cysteine synthesis both as a

  20. Characterization of a Leishmania isolate from the rodent host Neotoma micropus collected in Texas and comparison with human isolates.

    PubMed

    Grogl, M; Kreutzer, R D; McHugh, C P; Martin, R K

    1991-12-01

    We report the biological and biochemical parameters of Leishmania parasites (MNEO/US/90/WR972) isolated from a rodent host, Neotoma micropus, collected in Texas. Footpad inoculations of WR972 promastigotes into BALB/c mice and Syrian hamsters resulted in ulcerating lesions six and eight weeks post-inoculation, respectively. Using monoclonal antibody-stained touch preparations, amastigotes were found in the liver of both laboratory hosts. Infection of J774 macrophages with WR972 promastigotes supported the growth of amastigotes for 12 days at 35 degrees C. The WR972 parasite was identified by enzyme electrophoresis as L. mexicana. Isozyme comparison of WR972 with 42 L. mexicana isolates (from humans and rodents) from four different endemic areas, including Texas, suggest that these parasite populations are identical for approximately 97% of their genetic loci. Pulse field gel electrophoresis (PFGE) of WR972 resolved 18 chromosomes with a size range of 300- greater than 2,000 kb. The karyotype strongly resembles that of two other Texas L. mexicana isolates from humans. Taken together, the PFGE, hybridization, and isoenzyme data suggest that the wood rat isolate (WR972) is identical to parasites from human cutaneous lesions isolated in Texas and Central America. In addition, the biological characteristics of WR972, its infectivity of BALB/c mice and the Syrian hamster, and the potential of the isolate to infect, transform, and divide in J774 macrophages indicate that WR972 will be pathogenic in humans if transmission occurs. Health care providers should consider this possibility when studying the epidemiology and control of cutaneous leishmaniasis in Texas. PMID:1763798

  1. Characterization of phosphoglycan-containing secretory products of Leishmania.

    PubMed

    Ilg, T; Stierhof, Y D; Wiese, M; McConville, M J; Overath, P

    1994-01-01

    This article presents an overview on phosphoglycan-containing components secreted by the insect and mammalian stages of several species of Leishmania, the causative agents of leishmaniasis in the Old and New World. Firstly, promastigotes of all three species considered, L. mexicana, L. donovani and L. major, shed lipophosphoglycan (LPG) into the culture medium possibly by release of micelles from the cell surface. Like the cell-associated LPG, culture supernatant LPG is amphiphilic and composed of a lysoalkylphosphatidylinositol-phosphosaccharide core connected to species-specific phosphosaccharide repeats and oligosaccharide caps. Secondly, all three species release hydrophilic phosphoglycan. Thirdly, all three species appear to secrete proteins covalently modified by phosphosaccharide repeats and oligosaccharide caps. In the case of promastigotes of L. mexicana, these components are organized as two filamentous polymers released from the flagellar pocket: the secreted acid phosphatase (sAP) composed of a 100 kDa phosphoglycoprotein and a protein-containing high-molecular-weight-phosphoglycan (proteo-HMWPG) and fibrous networks likewise composed of phosphoglycan possibly linked to protein. Structural analyses and gene cloning suggest that the parasites can covalently modify protein regions rich in serine and threonine residues by the attachment of phosphosaccharide repeats capped by oligosaccharides. We propose that the networks formed in vitro correspond to fibrous material previously demonstrated in the digestive tract of infected sandflies. In the case of L. donovani, the sAP is also modified by phosphoglycans but contains neither proteo-HMWPG nor does it aggregate to filaments. Finally, L. mexicana amastigotes release proteo-HMWPG via the flagellar pocket into the parasitophorous vacuole of infected macrophages.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8084657

  2. Antimycobacterial and antileishmanial effects of microfungi isolated from tropical regions in México.

    PubMed

    Gamboa-Angulo, Marcela; Molina-Salinas, Gloria M; Chan-Bacab, Manuel; Peraza-Sánchez, Sergio R; Heredia, Gabriela; de la Rosa-García, Susana C; Reyes-Estebanez, Manuela

    2013-02-01

    A total of 82 fungal extracts were selected and screened against Mycobacterium tuberculosis and promastigotes of Leishmania mexicana strains. Results showed inhibitory activity in 29 % of the fungal strains against at least one of the targets tested. The most significant antituberculosis (antiTB) effects were presented by Cylindrocarpon sp. XH9B, Fusarium sp. TA54, Fusarium XH1Ga, Gliocladium penicillioides TH04 and TH21, Gliocladium sp. TH16, Kutilakesa sp. MR46, and Verticillium sp. TH28 strains (minimal inhibition concentration (MIC) = 1.56-25 μg/ml). Mortality of L. mexicana promastigotes was displayed by only four strains, Fusarium sp. TA50, Fusarium sp. TA54, Verticillium sp. TH28, and the unidentified 2TA2 strain (IC(50) = 14.23-100 μg/ml and IC(100) = 50-100 μg/ml). Seven of these most active strains were defatted and their corresponding fractions evaluated again. The results showed the best antiTB activity in Gliocladium sp. TH16 (MIC = 1.56 μg/ml) and the highest leishmanicidal potential in Fusarium sp. TA54 (IC(50) = 6.36 μg/ml). These results show that fungi living in the tropical regions of México have the ability to produce bioactive metabolites that could be used in the near future as natural products to control neglected tropical diseases. PMID:23086442

  3. Flagellar pocket restructuring through the Leishmania life cycle involves a discrete flagellum attachment zone.

    PubMed

    Wheeler, Richard J; Sunter, Jack D; Gull, Keith

    2016-02-15

    Leishmania promastigote parasites have a flagellum, which protrudes from the flagellar pocket at the cell anterior, yet, surprisingly, have homologs of many flagellum attachment zone (FAZ) proteins--proteins used in the related Trypanosoma species to laterally attach the flagellum to the cell body from the flagellar pocket to the cell posterior. Here, we use seven Leishmania mexicana cell lines that expressed eYFP fusions of FAZ protein homologs to show that the Leishmania flagellar pocket includes a FAZ structure. Electron tomography revealed a precisely defined 3D organisation for both the flagellar pocket and FAZ, with striking similarities to those of Trypanosoma brucei. Expression of two T. brucei FAZ proteins in L. mexicana showed that T. brucei FAZ proteins can assemble into the Leishmania FAZ structure. Leishmania therefore have a previously unrecognised FAZ structure, which we show undergoes major structural reorganisation in the transition from the promastigote (sandfly vector) to amastigote (in mammalian macrophages). Morphogenesis of the Leishmania flagellar pocket, a structure important for pathogenicity, is therefore intimately associated with a FAZ; a finding with implications for understanding shape changes involving component modules during evolution. PMID:26746239

  4. Flagellar pocket restructuring through the Leishmania life cycle involves a discrete flagellum attachment zone

    PubMed Central

    Wheeler, Richard J.; Sunter, Jack D.; Gull, Keith

    2016-01-01

    ABSTRACT Leishmania promastigote parasites have a flagellum, which protrudes from the flagellar pocket at the cell anterior, yet, surprisingly, have homologs of many flagellum attachment zone (FAZ) proteins – proteins used in the related Trypanosoma species to laterally attach the flagellum to the cell body from the flagellar pocket to the cell posterior. Here, we use seven Leishmania mexicana cell lines that expressed eYFP fusions of FAZ protein homologs to show that the Leishmania flagellar pocket includes a FAZ structure. Electron tomography revealed a precisely defined 3D organisation for both the flagellar pocket and FAZ, with striking similarities to those of Trypanosoma brucei. Expression of two T. brucei FAZ proteins in L. mexicana showed that T. brucei FAZ proteins can assemble into the Leishmania FAZ structure. Leishmania therefore have a previously unrecognised FAZ structure, which we show undergoes major structural reorganisation in the transition from the promastigote (sandfly vector) to amastigote (in mammalian macrophages). Morphogenesis of the Leishmania flagellar pocket, a structure important for pathogenicity, is therefore intimately associated with a FAZ; a finding with implications for understanding shape changes involving component modules during evolution. PMID:26746239

  5. Replication Attempt: “Effect of BMAP-28 Antimicrobial Peptides on Leishmania Major Promastigote and Amastigote Growth: Role of Leishmanolysin in Parasite Survival”

    PubMed Central

    Iorns, Elizabeth; Gunn, William; Erath, Jessey; Rodriguez, Ana; Zhou, Jian; Benzinou, Michael

    2014-01-01

    This study describes an attempt to replicate experiments from the paper “Effect of BMAP-28 Antimicrobial Peptides on Leishmania major Promastigote and Amastigote Growth: Role of Leishmanolysin in Parasite Survival,” which was submitted to the Reproducibility Initiative for independent validation. The cathelicidin bovine myeloid antimicrobial peptide 28 (BMAP-28) and its isomers were previously shown to have potent antiparasitic activity against Leishmania major. We tested the effectiveness of L-BMAP-28 and two of its isomers, the D-amino acid form (D-BMAP-28) and the retro-inverso form (RI-BMAP-28), in both unamidated and amidated forms, as anti-leishmanial agents against Leishmania major promastigotes in vitro. We observed that L-BMAP-28, as well as its D and RI isomers, demonstrate anti-leishmanial activity against L. major promastigotes in vitro. The inhibitory effect was lower than what was seen in the original study. At 2 µM of amidated peptides, the viability was 94%, 36%, and 66% with L-, D- and RI-peptides, versus 57%, 6%, and 18% in the original study. PMID:25517992

  6. Inhibition of Leishmania donovani promastigote DNA topoisomerase I and human monocyte DNA topoisomerases I and II by antimonial drugs and classical antitopoisomerase agents.

    PubMed

    Walker, John; Saravia, Nancy G

    2004-10-01

    We have compared the inhibitor sensitivities of DNA topoisomerase I (TOPI) from Leishmania donovani promastigotes and TOPs I and II of human monocytes using pentavalent and trivalent antimonials (SbV, SbIII) and classical TOP inhibitors. Bis-benzimidazoles (Hoechst-33258 and -33342) were potent inhibitors of both parasite and human TOPI, but Hoechst-33342 was markedly less cytotoxic to promastigotes than to monocytes in vitro. Leishmania donovani was also considerably less sensitive than monocytes to camptothecin, both at enzyme and cellular levels. Sodium stibogluconate (SSG) was the only antimonial to inhibit TOPI, exhibiting a significant (P < 0.05) 3-fold greater potency against the L. donovani enzyme but showed low cytotoxicities against intact promastigotes. The SbV meglumine antimoniate failed to inhibit TOPI and showed negligible cytotoxicities, whereas SbIII drugs were lethal to parasites and monocytes yet poor inhibitors of TOPI. Monocyte TOPII was inhibited by bis-benzimidazoles and insensitive to antimonials and camptothecin. The disparity between the high leishmanicidal activity and low anti-TOPI potency of SbIII indicates that in vivo targeting of L. donovani TOPI by the reductive pathway of antimonial activation is improbable. Nevertheless, the potent direct inhibition of TOPI by SSG and the differential interactions of camptothecin with L. donovani and human TOPI support the possibility of developing parasite-specific derivatives. PMID:15562618

  7. Exclusion of synaptotagmin V at the phagocytic cup by Leishmania donovani lipophosphoglycan results in decreased promastigote internalization.

    PubMed

    Vinet, Adrien F; Jananji, Silvana; Turco, Salvatore J; Fukuda, Mitsunori; Descoteaux, Albert

    2011-09-01

    Regulators of membrane fusion play an important role in phagocytosis, as they regulate the focal delivery of endomembrane that is required for optimal internalization of large particles. During internalization of Leishmania promastigotes, the surface glycolipid lipophosphoglycan (LPG) is transferred to the macrophage membrane and modifies its fusogenic properties. In this study, we investigated the impact of LPG on the recruitment of the exocytosis regulator synaptotagmin V (Syt V) at the area of internalization and on the early steps of phagocytosis. Using Leishmania donovani LPG-defective mutants and LPG-coated particles, we established that LPG reduces the phagocytic capacity of macrophages and showed that it causes exclusion of Syt V from the nascent phagosome. Silencing of Syt V inhibited phagocytosis to the same extent as LPG, and these effects were not cumulative, consistent with a Syt V-dependent mechanism for the inhibition of phagocytosis by LPG. Previous work has revealed that LPG-mediated exclusion of Syt V from phagosomes prevents the recruitment of the vacuolar ATPase and acidification. Thus, whereas exclusion of Syt V from phagosomes in the process of formation may be beneficial for the creation of a hospitable intracellular niche, it reduces the phagocytic capacity of macrophages. We propose that the cost associated with a reduced internalization rate may be compensated by increased survival, and could lead to a greater overall parasite fitness. PMID:21680635

  8. Cytochemical localization of ATP diphosphohydrolase from Leishmania (Viannia) braziliensis promastigotes and identification of an antigenic and catalytically active isoform.

    PubMed

    Rezende-Soares, F A; Carvalho-Campos, C; Marques, M J; Porcino, G N; Giarola, N L L; Costa, B L S; Taunay-Rodrigues, A; Faria-Pinto, P; Souza, M A; Diniz, V A; Corte-Real, S; Juliano, M A; Juliano, L; Vasconcelos, E G

    2010-04-01

    An ATP diphosphohydrolase (EC 3.6.1.5) activity was identified in a Leishmania (Viannia) braziliensis promastigotes preparation (Lb). Ultrastructural cytochemical microscopy showed this protein on the parasite surface and also stained a possible similar protein at the mitochondrial membrane. Isolation of an active ATP diphosphohydrolase isoform from Lb was obtained by cross-immunoreactivity with polyclonal anti-potato apyrase antibodies. These antibodies, immobilized on Protein A-Sepharose, immunoprecipitated a polypeptide of approximately 48 kDa and, in lower amount, a polypeptide of approximately 43 kDa, and depleted 83% ATPase and 87% of the ADPase activities from detergent-homogenized Lb. Potato apyrase was recognized in Western blots by IgG antibody from American cutaneous leishmaniasis (ACL) patients, suggesting that the parasite and vegetable proteins share antigenic conserved epitopes. Significant IgG seropositivity in serum samples diluted 1:50 from ACL patients (n=20) for Lb (65%) and potato apyrase (90%) was observed by ELISA technique. Significant IgG antibody reactivity was also observed against synthetic peptides belonging to a conserved domain from L. braziliensis NDPase (80% seropositivity) and its potato apyrase counterpart (50% seropositivity), in accordance with the existence of shared antigenic epitopes and demonstrating that in leishmaniasis infection the domain r82-103 from L. braziliensis NDPase is a target for the human immune response. PMID:19961654

  9. Different Host Complement Systems and Their Interactions with Saliva from Lutzomyia longipalpis (Diptera, Psychodidae) and Leishmania infantum Promastigotes

    PubMed Central

    Mendes-Sousa, Antonio Ferreira; Nascimento, Alexandre Alves Sousa; Queiroz, Daniel Costa; Vale, Vladimir Fazito; Fujiwara, Ricardo Toshio; Araújo, Ricardo Nascimento; Pereira, Marcos Horácio; Gontijo, Nelder Figueiredo

    2013-01-01

    Background Lutzomyia longipalpis is the vector of Leishmania infantum in the New World, and its saliva inhibits classical and alternative human complement system pathways. This inhibition is important in protecting the insect´s midgut from damage by the complement. L. longipalpis is a promiscuous blood feeder and must be protected against its host’s complement. The objective of this study was to investigate the action of salivary complement inhibitors on the sera of different host species, such as dogs, guinea pigs, rats and chickens, at a pH of 7.4 (normal blood pH) and 8.15 (the midgut pH immediately after a blood meal). We also investigated the role of the chicken complement system in Leishmania clearance in the presence and absence of vector saliva. Results The saliva was capable of inhibiting classical pathways in dogs, guinea pigs and rats at both pHs. The alternative pathway was not inhibited except in dogs at a pH of 8.15. The chicken classical pathway was inhibited only by high concentrations of saliva and it was better inhibited by the midgut contents of sand flies. Neither the saliva nor the midgut contents had any effect on the avian alternative pathway. Fowl sera killed L. infantum promastigotes, even at a low concentration (2%), and the addition of L. longipalpis saliva did not protect the parasites. The high body temperature of chickens (40°C) had no effect on Leishmania viability during our assays. Conclusion Salivary inhibitors act in a species-specific manner. It is important to determine their effects in the natural hosts of Leishmania infantum because they act on canid and rodent complements but not on chickens (which do not harbour the parasite). Moreover, we concluded that the avian complement system is the probable mechanism through which chickens eliminate Leishmania and that their high body temperature does not influence this parasite. PMID:24255715

  10. Apoptosis-like death in Leishmania donovani promastigotes induced by eugenol-rich oil of Syzygium aromaticum.

    PubMed

    Islamuddin, Mohammad; Sahal, Dinkar; Afrin, Farhat

    2014-01-01

    Leishmaniasis consists of a complex spectrum of infectious diseases with worldwide distribution of which visceral leishmaniasis or kala-azar caused by Leishmania donovani is the most devastating. In the absence of vaccines, chemotherapy remains the mainstay for the control of leishmaniasis. The drugs of choice are expensive and associated with multiple adverse side effects. Because of these limitations, the development of new antileishmanial compounds is imperative and plants offer prospects in this regard. The present work was conducted to study the antileishmanial potential of oil from Syzygium aromaticum flower buds (clove). The S. aromaticum oil was characterized by gas chromatography and GC-MS and eugenol as well as eugenyl acetate were found to be the most abundant compounds, composing 59.75 % and 29.24 %, respectively of the oil. Our findings have shown that eugenol-rich essential oil from S. aromaticum (EROSA) possesses significant activity against L. donovani, with 50 % inhibitory concentration of 21 ± 0.16 µg ml(-1) and 15.24 ± 0.14 µg ml(-1), respectively, against promastigotes and intracellular amastigotes. Alterations in cellular morphology and growth reversibility assay substantiated the leishmanicidal activity of EROSA. The leishmanicidal effect was mediated via apoptosis as confirmed by externalization of phosphatidylserine, DNA nicking by TdT-mediated dUTP nick-end labelling (TUNEL) assay, dyskinetoplastidy, cell cycle arrest at sub-G0-G1 phase, loss of mitochondrial membrane potential and reactive oxygen species generation. EROSA presented no adverse cytotoxic effects against murine macrophages even at 200 µg ml(-1). Our studies authenticate the promising antileishmanial activity of EROSA, which is mediated by programmed cell death, and, accordingly, EROSA may be a source of novel agents for the treatment of leishmaniasis. PMID:24161990

  11. Evaluation of photodynamic antimicrobial therapy (PACT) against promastigotes form of the Leishmania (Viannia) braziliensis: in vitro study

    NASA Astrophysics Data System (ADS)

    Barbosa, Artur F. S.; Sangiorgi, Bruno B.; Galdino, Suely L.; Pitta, Ivan R.; Barral Netto, Manoel; Correia, Neandder A.; Pinheiro, Antônio L. B.

    2012-03-01

    Leishmaniasis is a complex disease that affects more than 12 million people in 88 countries worldwide. Leishmania (Viannia) braziliensis is the most common species in the Americas and the most important causative agent of cutaneous and mucocutaneous leishmaniasis in Brazil. The therapeutic arsenal routinely employed to treat patients with leishmaniasis is limited and unsatisfactory. For cutaneous leishmaniasis, pentavalent antimonials are the first line therapeutic scheme recommended by the WHO. These compounds are highly toxic, poorly tolerated and their effectiveness highly variable. In this work, a technique with, so far, an unknown disadvantage is discussed. The aim of this study was to verify the effectiveness of PACT in vitro, as a new technique for the treatment of Leishmaniasis. For this, semiconductor laser (λ = 660nm, 40mW, 4.2J/cm2, CW) associated to phenothiazine's derivatives (5 and 10 μg/ml, TBO, Methylene Blue or Phenothiazine) on the promastigotes form of Leishmania braziliensis in a single session was used. Viability of the parasites was assessed in quadruplicates of each group. The samples were removed and analyzed in a hemocytometer 72h after PACT. We found an important decrease in the number of viable parasites on all treated groups in comparison to their controls. The results of present study showed significant percentage of lethality (above 95%) of the protocol. The 99.23% of lethality was achieved with 10 μg/ml of TBO. No lethality was seen on groups treated neither with laser nor with each compounds separately. The results are promising and indicative that the use of PACT may be a powerful treatment of leishmaniasis when compared to already available ones.

  12. In vitro study of the photodynamic antimicrobial therapy (PACT) against promastigotes form of the leishmania (viannia) braziliensis: in vitro study

    NASA Astrophysics Data System (ADS)

    Barbosa, Artur F. S.; Sangiorgi, Bruno B.; Galdino, Suely L.; Pitta, Ivan R.; Barral-Netto, Manoel; Pinheiro, Antônio L. B.

    2013-03-01

    Leishmaniasis, a protozoan parasitic disease that remains a major worldwide health problem with high endemicity in developing countries. Treatment of cutaneous Leishmaniasis (CL) should be decided by the clinical lesions, etiological species and its potential to develop into mucosal Leishmaniasis. High cost, systemic toxicity, and diminished efficacy due to development of parasite resistance are the serious drawbacks of current treatment options. Thus, identifying new, effective, and safer anti-leishmanial drug(s) is of paramount importance. The aim of this study was to verify the effectiveness of PACT in vitro, as a new technique for the treatment of Leishmaniasis. For this, semiconductor laser (λ = 660nm, 40mW, 8.4J/cm2, CW) associated to phenothiazine's derivatives (5 and 10 μg/ml, TBO, Methylene Blue or Phenothiazine) on the promastigotes form of Leishmania braziliensis in a single session was used. Viability of the parasites was assessed in quadruplicates of each group. The samples were removed and analyzed in a hemocytometer 72h after PACT. We found an important decrease in the number of viable parasites on all treated groups in comparison to their controls. The results of present study showed significant percentage of lethality (above 92%) of the protocol. The 98.33% of lethality was achieved with 10 μg/ml of FTZ. No lethality was seen on groups treated neither with laser nor with each compounds separately. The results are promising and indicative that the use of PACT may be a powerful treatment of Leishmaniasis when compared to already available ones.

  13. Evaluation of Organelle Changes in Promastigotes of Unresponsive Leishmania Tropica to Meglumine Antimoniate in Comparison with Sensitive and Standard Isolates by Electron Microscopy

    PubMed Central

    Bahreini, Mitra; Bolorizadeh, Mehdi; Dabiri, Shahriar; Sharifi, Iraj

    2015-01-01

    Background: The control of leishmaniasis faces serious challenges because of resistance to the first-line antimonial drugs. We aimed to evaluate the differences in organelle changes of cultivated promastigotes obtained from skin lesions of sensitive and unresponsive isolates to meglumine antimoniate (Glucantime) by electron microscopy. Material and Methods: This study was done in Bam city, southeastern Iran, in which the incidence of disease has sharply increased since the earthquake in 2003. The samples were taken from 66 patients who were referred to the cutaneous leishmaniasis (CL) treatment center in Bam. A questionnaire was completed for each individual, recording their demographic characteristics and CL status. The scraping smears provided from the edge of active lesions with sterile blades were fixed with methanol, stained by Giemsa, and examined under a compound light microscope for amastigote form simultaneously. To prepare the specimens for transmission electron imaging, promastigotes were centrifuged and resuspened. Results: Transmission electron microscopic study of the cultivated promastigotes revealed that there were alterations in the organelles and structures of sensitive isolates compared with unresponsive and standard ones. Organelles and structures such as mitochondria, kinetoplast, microtubules, cytoplasmic vacuoles, plasma membrane and vesicles were studied. The alterations such as disintegration of kinetoplast into thin filaments and condensation of kinetoplast DNA core, changes in size, number and location of vesicles and microtubules were observed. We noted intense cytoplasmic vacuolization, and considerable swelling of mitochondria. Conclusion: The significance and relevance of these changes might help understand drug resistance patterns and help localize the best target site for inactivating the organism. PMID:26120175

  14. LdARL-3A, a Leishmania promastigote-specific ADP-ribosylation factor-like protein, is essential for flagellum integrity.

    PubMed

    Cuvillier, A; Redon, F; Antoine, J C; Chardin, P; DeVos, T; Merlin, G

    2000-06-01

    The small G protein-encoding LdARL-3A gene, a homologue of the human ARL-3 gene, was isolated from Leishmania donovani, and its protein product characterised. It is unique in the Leishmania genome and expressed only in the extracellular promastigote insect form, but not in the intracellular amastigote mammalian form, as shown by northern blots and western blots developed with a specific anti-C terminus immune serum. Indirect immunofluorescence microscopy revealed distinct labelled spots regularly distributed on the plasma membrane, including the part lining the flagellum and the flagellar pocket. By transfection experiments, it was found that wild-type LdARL-3A-overexpressing promastigotes reached higher densities in culture, but released significantly less secreted acid phosphatase in the extracellular medium than the parental strain. When LdARL-3A blocked under the GDP-bound 'inactive' form or with an inactivated potential myristoylation site was overexpressed, the cells displayed an apparent wild-type phenotype, but died earlier in the stationary phase; in contrast to parental cells, they showed a diffuse pattern of fluorescence labelling in the cytoplasm and on the cell membrane. Strikingly, when a constitutively 'active' form of LdARL-3A (blocked under the GTP-bound form) was overexpressed, the promastigotes were immobile with a very short flagellum, a slow growth rate and a low level of acid phosphatase secretion; the length of the flagellum was inversely proportional to mutant protein expression. We concluded that LdARL-3A could be an essential gene involved in flagellum biogenesis; it may provide new approaches for control of the parasite at the insect stage. PMID:10806117

  15. In Vitro Evaluation of a Soluble Leishmania Promastigote Surface Antigen as a Potential Vaccine Candidate against Human Leishmaniasis

    PubMed Central

    Bahi-Jaber, Narges; Petitdidier, Elodie; Markikou-Ouni, Wafa; Aoun, Karim; Moreno, Javier; Carrillo, Eugenia; Salotra, Poonam; Kaushal, Himanshu; Negi, Narender Singh; Arevalo, Jorge; Falconi-Agapito, Francesca; Privat, Angela; Cruz, Maria; Pagniez, Julie; Papierok, Gérard-Marie; Rhouma, Faten Bel Haj; Torres, Pilar; Lemesre, Jean-Loup; Chenik, Mehdi; Meddeb-Garnaoui, Amel

    2014-01-01

    PSA (Promastigote Surface Antigen) belongs to a family of membrane-bound and secreted proteins present in several Leishmania (L.) species. PSA is recognized by human Th1 cells and provides a high degree of protection in vaccinated mice. We evaluated humoral and cellular immune responses induced by a L. amazonensis PSA protein (LaPSA-38S) produced in a L. tarentolae expression system. This was done in individuals cured of cutaneous leishmaniasis due to L. major (CCLm) or L. braziliensis (CCLb) or visceral leishmaniasis due to L. donovani (CVLd) and in healthy individuals. Healthy individuals were subdivided into immune (HHR-Lm and HHR-Li: Healthy High Responders living in an endemic area for L. major or L. infantum infection) or non immune/naive individuals (HLR: Healthy Low Responders), depending on whether they produce high or low levels of IFN-γ in response to Leishmania soluble antigen. Low levels of total IgG antibodies to LaPSA-38S were detected in sera from the studied groups. Interestingly, LaPSA-38S induced specific and significant levels of IFN-γ, granzyme B and IL-10 in CCLm, HHR-Lm and HHR-Li groups, with HHR-Li group producing TNF-α in more. No significant cytokine response was observed in individuals immune to L. braziliensis or L. donovani infection. Phenotypic analysis showed a significant increase in CD4+ T cells producing IFN-γ after LaPSA-38S stimulation, in CCLm. A high positive correlation was observed between the percentage of IFN-γ-producing CD4+ T cells and the released IFN-γ. We showed that the LaPSA-38S protein was able to induce a mixed Th1 and Th2/Treg cytokine response in individuals with immunity to L. major or L. infantum infection indicating that it may be exploited as a vaccine candidate. We also showed, to our knowledge for the first time, the capacity of Leishmania PSA protein to induce granzyme B production in humans with immunity to L. major and L. infantum infection. PMID:24786587

  16. In vitro evaluation of a soluble Leishmania promastigote surface antigen as a potential vaccine candidate against human leishmaniasis.

    PubMed

    Chamakh-Ayari, Rym; Bras-Gonçalves, Rachel; Bahi-Jaber, Narges; Petitdidier, Elodie; Markikou-Ouni, Wafa; Aoun, Karim; Moreno, Javier; Carrillo, Eugenia; Salotra, Poonam; Kaushal, Himanshu; Negi, Narender Singh; Arevalo, Jorge; Falconi-Agapito, Francesca; Privat, Angela; Cruz, Maria; Pagniez, Julie; Papierok, Gérard-Marie; Rhouma, Faten Bel Haj; Torres, Pilar; Lemesre, Jean-Loup; Chenik, Mehdi; Meddeb-Garnaoui, Amel

    2014-01-01

    PSA (Promastigote Surface Antigen) belongs to a family of membrane-bound and secreted proteins present in several Leishmania (L.) species. PSA is recognized by human Th1 cells and provides a high degree of protection in vaccinated mice. We evaluated humoral and cellular immune responses induced by a L. amazonensis PSA protein (LaPSA-38S) produced in a L. tarentolae expression system. This was done in individuals cured of cutaneous leishmaniasis due to L. major (CCLm) or L. braziliensis (CCLb) or visceral leishmaniasis due to L. donovani (CVLd) and in healthy individuals. Healthy individuals were subdivided into immune (HHR-Lm and HHR-Li: Healthy High Responders living in an endemic area for L. major or L. infantum infection) or non immune/naive individuals (HLR: Healthy Low Responders), depending on whether they produce high or low levels of IFN-γ in response to Leishmania soluble antigen. Low levels of total IgG antibodies to LaPSA-38S were detected in sera from the studied groups. Interestingly, LaPSA-38S induced specific and significant levels of IFN-γ, granzyme B and IL-10 in CCLm, HHR-Lm and HHR-Li groups, with HHR-Li group producing TNF-α in more. No significant cytokine response was observed in individuals immune to L. braziliensis or L. donovani infection. Phenotypic analysis showed a significant increase in CD4+ T cells producing IFN-γ after LaPSA-38S stimulation, in CCLm. A high positive correlation was observed between the percentage of IFN-γ-producing CD4+ T cells and the released IFN-γ. We showed that the LaPSA-38S protein was able to induce a mixed Th1 and Th2/Treg cytokine response in individuals with immunity to L. major or L. infantum infection indicating that it may be exploited as a vaccine candidate. We also showed, to our knowledge for the first time, the capacity of Leishmania PSA protein to induce granzyme B production in humans with immunity to L. major and L. infantum infection. PMID:24786587

  17. Studies on the protective efficacy of freeze thawed promastigote antigen of Leishmania donovani along with various adjuvants against visceral leishmaniasis infection in mice.

    PubMed

    Thakur, Ankita; Kaur, Harpreet; Kaur, Sukhbir

    2015-09-01

    Visceral leishmaniasis (VL) caused by Leishmania donovani persists as a major public health issue in tropical and subtropical areas of the world. Current treatment of this disease relies on use of drugs. It is doubtful that chemotherapy can alone eradicate the disease, so there is a need for an effective vaccine. Killed antigen candidates remain a good prospect considering their ease of formulation, stability, low cost and safety. To enhance the efficacy of killed vaccines suitable adjuvant and delivery system are needed. Therefore, the current study was conducted to determine the protective efficacy of freeze-thawed L. donovani antigen in combination with different adjuvants against experimental infection of VL. For this, BALB/c mice were immunized thrice at an interval of two weeks. Challenge infection was given two weeks after last immunization. Mice were sacrificed after last immunization and on different post challenge/infection days. Immunized mice showed significant reduction in parasite burden, enhanced DTH responses with increased levels of Th1 cytokines and lower levels of Th2 cytokines, thus indicating the development of a protective Th1 response. Maximum protection was achieved with liposome encapsulated freeze thawed promastigote (FTP) antigen of L. donovani and it was followed by group immunized with FTP+MPL-A, FTP+saponin, FTP+alum and FTP antigen (alone). The present study highlights greater efficacy of freeze thawed promastigote antigen as a potential vaccine candidate along with effective adjuvant formulations against experimental VL infection. PMID:26001730

  18. Contrasting human cytokine responses to promastigote whole-cell extract and the Leishmania analogue receptor for activated C kinase antigen of L. amazonensis in natural infection versus immunization

    PubMed Central

    Azeredo-Coutinho, R B G; Matos, D C S; Armôa, G G R; Maia, R M; Schubach, A; Mayrink, W; Mendonça, S C F

    2008-01-01

    It is known that the same antigen can induce different immune responses, depending upon the way that it is presented to the immune system. The objective of this study was to compare cytokine responses of peripheral blood mononuclear cells (PBMC) from cutaneous leishmaniasis patients and subjects immunized with a first-generation candidate vaccine composed of killed Leishmania amazonensis promastigotes to a whole-cell promastigote antigen extract (La) and to the recombinant protein LACK (Leishmania analogue receptor for activated C kinase), both from L. amazonensis. Thirty-two patients, 35 vaccinees and 13 healthy subjects without exposure to Leishmania, were studied. Cytokine production was assessed by enzyme-linked immunosorbent assay and enzyme-linked immunospot assay. The interferon (IFN)-γ levels stimulated by La were significantly higher and the levels of interleukin (IL)-10 significantly lower than those stimulated by LACK in the patient group, while LACK induced a significantly higher IFN-γ production and a significantly lower IL-10 production compared with those induced by La in the vaccinated group. LACK also induced a significantly higher frequency of IFN-γ-producing cells than did La in the vaccinated group. The contrast in the cytokine responses stimulated by LACK and La in PBMC cultures from vaccinated subjects versus patients indicates that the human immune response to crude and defined Leishmania antigens as a consequence of immunization differs from that induced by natural infection. PMID:18627399

  19. Serum Removal from Culture Induces Growth Arrest, Ploidy Alteration, Decrease in Infectivity and Differential Expression of Crucial Genes in Leishmania infantum Promastigotes.

    PubMed

    Alcolea, Pedro J; Alonso, Ana; Moreno-Izquierdo, Miguel A; Degayón, María A; Moreno, Inmaculada; Larraga, Vicente

    2016-01-01

    Leishmania infantum is one of the species responsible for visceral leishmaniasis. This species is distributed basically in the Mediterranean basin. A recent outbreak in humans has been reported in Spain. Axenic cultures are performed for most procedures with Leishmania spp. promastigotes. This model is stable and reproducible and mimics the conditions of the gut of the sand fly host, which is the natural environment of promastigote development. Culture media are undefined because they contain mammalian serum, which is a rich source of complex lipids and proteins. Serum deprivation slows down the growth kinetics and therefore, yield in biomass. In fact, we have confirmed that the growth rate decreases, as well as infectivity. Ploidy is also affected. Regarding the transcriptome, a high-throughput approach has revealed a low differential expression rate but important differentially regulated genes. The most remarkable profiles are: up-regulation of the GINS Psf3, the fatty acyl-CoA synthase (FAS1), the glyoxylase I (GLO1), the hydrophilic surface protein B (HASPB), the methylmalonyl-CoA epimerase (MMCE) and an amastin gene; and down-regulation of the gPEPCK and the arginase. Implications for metabolic adaptations, differentiation and infectivity are discussed herein. PMID:26959417

  20. Serum Removal from Culture Induces Growth Arrest, Ploidy Alteration, Decrease in Infectivity and Differential Expression of Crucial Genes in Leishmania infantum Promastigotes

    PubMed Central

    Alcolea, Pedro J.; Alonso, Ana; Moreno-Izquierdo, Miguel A.; Degayón, María A.; Moreno, Inmaculada; Larraga, Vicente

    2016-01-01

    Leishmania infantum is one of the species responsible for visceral leishmaniasis. This species is distributed basically in the Mediterranean basin. A recent outbreak in humans has been reported in Spain. Axenic cultures are performed for most procedures with Leishmania spp. promastigotes. This model is stable and reproducible and mimics the conditions of the gut of the sand fly host, which is the natural environment of promastigote development. Culture media are undefined because they contain mammalian serum, which is a rich source of complex lipids and proteins. Serum deprivation slows down the growth kinetics and therefore, yield in biomass. In fact, we have confirmed that the growth rate decreases, as well as infectivity. Ploidy is also affected. Regarding the transcriptome, a high-throughput approach has revealed a low differential expression rate but important differentially regulated genes. The most remarkable profiles are: up-regulation of the GINS Psf3, the fatty acyl-CoA synthase (FAS1), the glyoxylase I (GLO1), the hydrophilic surface protein B (HASPB), the methylmalonyl-CoA epimerase (MMCE) and an amastin gene; and down-regulation of the gPEPCK and the arginase. Implications for metabolic adaptations, differentiation and infectivity are discussed herein. PMID:26959417

  1. Effect of glucantime on field and patient isolates of New World Leishmania: use of growth parameters of promastigotes to assess antimony susceptibility.

    PubMed

    Moreira, E S; Anacleto, C; Petrillo-Peixoto, M L

    1998-09-01

    The effect of pentavalent meglumine antimoniate (glucantime) on the growth kinetics of promastigotes of 13 South American Leishmania strains isolated from patients, sylvatic reservoirs, and vectors was studied. Four of five L. (Viannia) braziliensis human isolates were obtained from drug-responsive patients and one was isolated from an unresponsive mucocutaneous-type infection. Studies involved the cell yield at the late log phase, the growth rate, and the growth-curve patterns of promastigotes in vitro. Restriction-fragment-length polymorphism, pulsed-field gel electrophoresis, and hybridization with the gene coding for a P-glycoprotein from L. (V.) guyanensis were used in an attempt to correlate the resistance phenotype with gene amplification. Consistent differences observed in both cell yield and growth rate among the isolates in the presence of glucantime indicated these parameters to be good criteria for the estimation of susceptibility to glucantime. Drug susceptibility varied widely between strains, indicating a great genetic heterogeneity of the isolates. Five L. (V.) braziliensis strains and three L. (V.) guyanensis strains were shown to be susceptible to glucantime. Five isolates were resistant, four of which were obtained from sylvatic vectors and one, from a patient with an unresponsive mucocutaneous infection. Molecular analyses of total DNA indicated the presence of a pgpA-related gene in all strains tested. No amplified sequence could be detected, suggesting that pgpA amplification is not involved in glucantime resistance in these wild Leishmania strains. PMID:9766900

  2. Nivel de Actividad Física, Autoeficacia, Beneficios y Barreras Percibidas en Mujeres Mayores Mexicanas Independientes (Physical Activity Level, Exercise Self-Efficacy, Benefits and Perceived Barriers of Independent Mexican Older Women).

    PubMed

    Enríquez-Reyna, María Cristina; Cruz-Castruita, Rosa María; Zamarripa, Jorge; Ceballos-Gurrola, Oswaldo; Guevara-Valtier, Milton Carlos

    2016-03-01

    This descriptive comparative study examined differences in personal characteristics, exercise self-efficacy, benefits and barriers of independent elderly women to perform physical activity (PA) according with the PA level. Two hundred three women older than 60 years of age, from a community located in Nuevo Leon, Mexico participated in the study. Data was collected using: a) A personal data questionnaire, b) Exercise Self-Efficacy Scale, c) Exercise Benefits/ Barriers Scale and d) Physical Activity Scale for the Elderly were completed. Age was similar in participants with low and acceptable PA level. Participants with lower levels of PA reported consuming more medications, fewer years of education and lower values of exercise self-efficacy, benefits and barriers. In this sample, exercise self-efficacy and benefits were positively associated with the PA level. PMID:27257188

  3. Survival in an extreme habitat: the roles of behaviour and energy limitation.

    PubMed

    Plath, Martin; Tobler, Michael; Riesch, Rüdiger; García de León, Francisco J; Giere, Olav; Schlupp, Ingo

    2007-12-01

    Extreme habitats challenge animals with highly adverse conditions, like extreme temperatures or toxic substances. In this paper, we report of a fish (Poecilia mexicana) inhabiting a limestone cave in Mexico. Several springs inside the cave are rich in toxic H(2)S. We demonstrate that a behavioural adaptation, aquatic surface respiration (ASR), allows for the survival of P. mexicana in this extreme, sulphidic habitat. Without the possibility to perform ASR, the survival rate of P. mexicana was low even at comparatively low H(2)S concentrations. Furthermore, we show that food limitation affects the survival of P. mexicana pointing to energetically costly physiological adaptations to detoxify H(2)S. PMID:17639290

  4. Survival in an extreme habitat: the roles of behaviour and energy limitation

    NASA Astrophysics Data System (ADS)

    Plath, Martin; Tobler, Michael; Riesch, Rüdiger; García de León, Francisco J.; Giere, Olav; Schlupp, Ingo

    2007-12-01

    Extreme habitats challenge animals with highly adverse conditions, like extreme temperatures or toxic substances. In this paper, we report of a fish ( Poecilia mexicana) inhabiting a limestone cave in Mexico. Several springs inside the cave are rich in toxic H2S. We demonstrate that a behavioural adaptation, aquatic surface respiration (ASR), allows for the survival of P. mexicana in this extreme, sulphidic habitat. Without the possibility to perform ASR, the survival rate of P. mexicana was low even at comparatively low H2S concentrations. Furthermore, we show that food limitation affects the survival of P. mexicana pointing to energetically costly physiological adaptations to detoxify H2S.

  5. Sterols with antileishmanial activity isolated from the roots of Pentalinon andrieuxii

    PubMed Central

    Pan, Li; Lezama-Davila, Claudio M.; Isaac-Marquez, Angelica P.; Calomeni, Edward P.; Fuchs, James R.; Satoskar, Abhay R.; Kinghorn, A. Douglas

    2012-01-01

    A new cholesterol derivative, pentalinonsterol (cholest-4,20,24-trien-3-one, 1), and a new polyoxygenated pregnane sterol glycoside, pentalinonside (2), together with 18 known compounds, including 14 sterols (3–16), three coumarins (17–19), and a triterpene (20), were isolated from a n-hexane partition of a methanol extract of the roots of the Mexican medicinal plant Pentalinon andrieuxii. Structure elucidation of compounds 1 and 2 was accomplished by spectroscopic data interpretation. All isolates were evaluated in vitro for their antileishmanial activity. Among these compounds, 6,7-dihydroneridienone (15) was found to be the most potent principle against promastigotes of Leishmania mexicana (L. mexicana). The cholesterol analogue, pentalinonsterol (1), together with two known sterols, 24-methylcholest-4,24(28)-dien-3-one (3) and neridienone (16), also exhibited significant leishmanicidal activity in this same bioassay. Compounds 1, 3, 15, 16, cholest-4-en-3-one (4), and cholest-5,20,24-trien-3β-ol (7), showed strong antileishmanial activity against amastigotes of L. mexicana, and 4 was found to be the most potent agent with an IC50 value of 0.03 μM. All the isolates were also evaluated for their cytotoxicity in non-infected bone marrow-derived macrophages, but none of these compounds was found active towards this cell line. The intracellular parasites treated with compounds 1, 3, 4, 15, and 16 were further studied by electron microscopy; morphological abnormalities and destruction of the amastigotes were observed, as a result of treatment with these compounds. PMID:22840389

  6. Identification and characterization of new Leishmania promastigote surface antigens, LaPSA-38S and LiPSA-50S, as major immunodominant excreted/secreted components of L. amazonensis and L. infantum.

    PubMed

    Bras-Gonçalves, Rachel; Petitdidier, Elodie; Pagniez, Julie; Veyrier, Renaud; Cibrelus, Prisca; Cavaleyra, Mireille; Maquaire, Sarah; Moreaux, Jérôme; Lemesre, Jean-Loup

    2014-06-01

    We have previously demonstrated that sera from dogs vaccinated with excreted/secreted antigens (ESA) of Leishmania infantum promastigotes (LiESAp) mainly recognized an immunodominant antigen of 54 kDa. An anti-LiESAp-specific IgG2 humoral response was observed and associated to Th1-type response in vaccinated dogs. This response was highly correlated with a long-lasting and strong LiESAp-vaccine protection toward L. infantum experimental infection. In addition, it was also shown that dogs from the vaccinated group developed a selective IgG2 response against an immunodominant antigen of 45 kDa of Leishmania amazonensis ESA promastigotes (LaESAp). In order to identify and characterize these immunodominant antigens, a mouse monoclonal antibody (mAb F5) was produced by immunization against LaESAp. It was found to recognize the major antigenic targets of both LaESAp and LiESAp. Analysis with mAb F5 of L. amazonensis amastigote and promastigote cDNA expression libraries enabled the identification of clones encoding proteins with significant structural homology to the promastigote surface antigens named PSA-2/gp-46. Among them, one clone presented a full-length cDNA and encoded a novel L. amazonensis protein of 38.6 kDa calculated molecular mass (LaPSA-38S) sharing an amino acid sequence consistent with that of the PSA polymorphic family and a N-terminal signal peptide, characteristic of a secreted protein. We then screened a L. infantum promastigote DNA cosmid library using a cDNA probe derived from the LaPSA-38S gene and identified a full-length clone of a novel excreted/secreted protein of L. infantum with a calculated molecular mass of 49.2 kDa and named LiPSA-50S. The fact that a significant immunological reactivity was observed against PSA, suggests that these newly identified proteins could have an important immunoregulatory influence on the immune response. This hypothesis is supported by the fact that (i) these proteins were naturally excreted/secreted by viable

  7. Interacting Protein Kinases Involved in the Regulation of Flagellar Length

    PubMed Central

    Erdmann, Maja; Scholz, Anne; Melzer, Inga M.; Schmetz, Christel; Wiese, Martin

    2006-01-01

    A striking difference of the life stages of the protozoan parasite Leishmania is a long flagellum in the insect stage promastigotes and a rudimentary organelle in the mammalian amastigotes. LmxMKK, a mitogen-activated protein (MAP) kinase kinase from Leishmania mexicana, is required for growth of a full-length flagellum. We identified LmxMPK3, a MAP kinase homologue, with a similar expression pattern as LmxMKK being not detectable in amastigotes, up-regulated during the differentiation to promastigotes, constantly expressed in promastigotes, and shut down during the differentiation to amastigotes. LmxMPK3 null mutants resemble the LmxMKK knockouts with flagella reduced to one-fifth of the wild-type length, stumpy cell bodies, and vesicles and membrane fragments in the flagellar pocket. A constitutively activated recombinant LmxMKK activates LmxMPK3 in vitro. Moreover, LmxMKK is likely to be directly involved in the phosphorylation of LmxMPK3 in vivo. Finally, LmxMPK3 is able to phosphorylate LmxMKK, indicating a possible feedback regulation. This is the first time that two interacting components of a signaling cascade have been described in the genus Leishmania. Moreover, we set the stage for the analysis of reversible phosphorylation in flagellar morphogenesis. PMID:16467378

  8. Rapid identification of Leishmania species by specific hybridization of kinetoplast DNA in cutaneous lesions.

    PubMed Central

    Wirth, D F; Pratt, D M

    1982-01-01

    Kinetoplast DNA (kDNA) was isolated from various species of the protozoic parasite Leishmania and analyzed by nucleic acid hybridization to detect species-related heterogeneity of kDNA. Purified DNA isolated from L. mexicana and L. braziliensis displayed no homology in nucleic acid hybridization studies. These results confirmed that rapid kDNA sequence change and evolution is occurring in New World species of Leishmania and suggested that such isolated kDNA could be used as a specific hybridization probe for the rapid identification of Leishmania species by using whole organisms. This work further demonstrates that such species-specific identification is feasible on isolated Leishmania promastigotes and, more important, directly on tissue touch blots derived from the cutaneous lesion. Thus, specific hybridization of isolated kDNA provides the basis for a rapid, accurate method for the diagnosis of human leishmaniasis directly from infected tissue. Images PMID:6960359

  9. Rapid identification of Leishmania species by specific hybridization of kinetoplast DNA in cutaneous lesions.

    PubMed

    Wirth, D F; Pratt, D M

    1982-11-01

    Kinetoplast DNA (kDNA) was isolated from various species of the protozoic parasite Leishmania and analyzed by nucleic acid hybridization to detect species-related heterogeneity of kDNA. Purified DNA isolated from L. mexicana and L. braziliensis displayed no homology in nucleic acid hybridization studies. These results confirmed that rapid kDNA sequence change and evolution is occurring in New World species of Leishmania and suggested that such isolated kDNA could be used as a specific hybridization probe for the rapid identification of Leishmania species by using whole organisms. This work further demonstrates that such species-specific identification is feasible on isolated Leishmania promastigotes and, more important, directly on tissue touch blots derived from the cutaneous lesion. Thus, specific hybridization of isolated kDNA provides the basis for a rapid, accurate method for the diagnosis of human leishmaniasis directly from infected tissue. PMID:6960359

  10. Biochemical mechanisms of the antileishmanial activity of sodium stibogluconate.

    PubMed Central

    Berman, J D; Waddell, D; Hanson, B D

    1985-01-01

    Pentavalent antimonial agents such as sodium stibogluconate (Pentostam; Burroughs Wellcome Co., London, United Kingdom) are the drugs of choice for the treatment of leishmaniasis, but their biochemical mechanisms of action are virtually unknown. The viability of Leishmania mexicana (WR 227) promastigotes and amastigotes was decreased 40 to 61% by a 4-h exposure to 500 micrograms of Sb (in the form of stibogluconate) per ml. Such exposure also resulted in a 51 to 65% decrease in incorporation of label into DNA, RNA, and protein; a 56 to 65% decrease in incorporation of label into purine nucleoside triphosphate; and a 34 to 60% increase in incorporation of label into purine nucleoside monophosphate and diphosphate. It is postulated that the apparent decrease in ATP and GTP synthesis from ADP and GDP contributes to decreased macromolecular synthesis and to decreased Leishmania viability. Further experiments suggested that inhibition of glycolysis and the citric acid cycle may partially explain the inability to phosphorylate ADP. PMID:2411217

  11. Prophylactic immunization against experimental leishmaniasis. III. Protection against fatal Leishmania tropica infection induced by irradiated promastigotes involves Lyt-1/sup +/2/sup -/ T cells that do not mediate cutaneous DTH

    SciTech Connect

    Liew, F.Y.; Howard, J.G.; Hale, C.

    1984-01-01

    Protective immunity against fatal L. tropica infection in genetically vulnerable BALB/c mice can be induced by prophylactic immunization with irradiated promastigotes even when heat-killed. Such immunity is adoptively transferable transiently into intact or durably into sub-lethally irradiated (200 or 550 rad) syngeneic recipients by splenic T but not B cells. The effector T cells are of the Lyt-1/sup +/2/sup -/ phenotype, devoid of demonstrable cytotoxic activity. The immune splenic T cell population expresses specific helper activity for antibody synthesis. A causal role for helper T cells in this capacity, however, seems unlikely, because it was shown that antibody does not determine the protective immunity against L. tropica. The immunized donors show no detectable cutaneous DTH or its early memory recall in response to live or killed promastigotes or a soluble L. tropica antigen preparation. Spleen, lymph node, and peritoneal exudate cells from protectively immunized donors similarly fail to transfer DTH locally or systemically. These cells also lack demonstrable suppressive activity against the expression or induction of DTH to L. tropica. Thus, protection against L. tropica induced by prophylactic i.v. immunization with irradiated promastigotes appears to be conferred by Lyt-1/sup +/2/sup -/ T cells that are distinguishable from T cells mediating either both DTH and T help, or cytotoxicity.

  12. Phototoxic effects of silicon bis (dimetilaminoetanoxi)-phthalocyanine (SiPc) on the viability of Leishmania major and Leishmania braziliensis promastigotes

    NASA Astrophysics Data System (ADS)

    Guerra Pinto, Juliana; Ferreira-Strixino, Juliana; Mittmann, Josane

    2016-06-01

    American cutaneous leishmaniasis (ACL) is an infectious disease caused by protozoans of the genus Leishmania. The treatment may consist of pentavalent antimonials or pentamidine and amphotericin. However, these treatments are extremely aggressive. Photodynamic antimicrobial chemotherapy (PACT) involves the same mechanism of photodynamic therapy which associates a photosensitizer with oxygen and a light source generating a photochemical reaction leading to cell death. The aim of this study was to verify the potential use of silicon bis (dimetilaminoetanoxi)-phthalocyanine (SiPc) compound in photodynamic treatment through evaluation of its phototoxic effect in promastigotes of the genus Leishmania braziliensis and Leishmania major. Treatment with SiPc was able to drastically affect the viability of the parasites as well as affect their growth and morphology, after PACT treatment. The data shown in this study allows us to conclude that SiPc is a promising photosensitizer (PS) since it does not affect parasite growth and viability in the dark. After PACT with this phthalocyanine, over 99% of parasites were killed with the higher concentration and a light dose used. These results suggest that SiPc can be used in future to treat CL, however, further studies are necessary to determine whether the PS are toxic to mononuclear phagocytic cells and epithelial cells which will also be affected by therapy when applied topically.

  13. The Leishmania donovani UMP Synthase Is Essential for Promastigote Viability and Has an Unusual Tetrameric Structure That Exhibits Substrate-controlled Oligomerization

    SciTech Connect

    French, Jarrod B.; Yates, Phillip A.; Soysa, D.Radika; Boitz, Jan M.; Carter, Nicola S.; Chang, Bailey; Ullman, Buddy; Ealick, Steven E.

    2011-08-09

    The final two steps of de novo uridine 5'-monophosphate (UMP) biosynthesis are catalyzed by orotate phosphoribosyltransferase (OPRT) and orotidine 5'-monophosphate decarboxylase (OMPDC). In most prokaryotes and simple eukaryotes these two enzymes are encoded by separate genes, whereas in mammals they are expressed as a bifunctional gene product called UMP synthase (UMPS), with OPRT at the N terminus and OMPDC at the C terminus. Leishmania and some closely related organisms also express a bifunctional enzyme for these two steps, but the domain order is reversed relative to mammalian UMPS. In this work we demonstrate that L. donovani UMPS (LdUMPS) is an essential enzyme in promastigotes and that it is sequestered in the parasite glycosome. We also present the crystal structure of the LdUMPS in complex with its product, UMP. This structure reveals an unusual tetramer with two head to head and two tail to tail interactions, resulting in two dimeric OMPDC and two dimeric OPRT functional domains. In addition, we provide structural and biochemical evidence that oligomerization of LdUMPS is controlled by product binding at the OPRT active site. We propose a model for the assembly of the catalytically relevant LdUMPS tetramer and discuss the implications for the structure of mammalian UMPS.

  14. IMP Dehydrogenase Deficiency in Leishmania donovani Causes a Restrictive Growth Phenotype in Promastigotes but is not Essential for Infection in Mice

    PubMed Central

    Fulwiler, Audrey L.; Boitz, Jan. M.; Gilroy, Caslin; Yates, Phillip A.; Jardim, Armando; Ullman, Buddy

    2011-01-01

    Leishmania cannot synthesize purines de novo and therefore must scavenge purines from its host for survival and growth. Biochemical and genomic analyses have indicated that Leishmania species express three potential routes for the synthesis of guanylate nucleotides: 1) a two-step pathway that converts IMP to GMP; 2) a three-step pathway that starts with the deamination of guanine to xanthine, followed by phosphoribosylation to XMP and then conversion to GMP; or 3) direct guanine phosphoribosylation by HGPRT. To determine the role of the first of these pathways to guanylate nucleotide synthesis, an L. donovani line deficient in IMP dehydrogenase (IMPDH), the first step in the IMP to GMP pathway, was constructed by targeted gene replacement. The Δimpdh lesion triggered a highly restrictive growth phenotype in promastigotes in culture but did not impact parasitemias in mice. The dispensability of IMPDH in vivo is the first definitive demonstration that intracellular L. donovani amastigotes have access to a sufficient pool of guanine, xanthine, or guanylate precursors from the host. PMID:21907738

  15. Proteomic Approach for Characterization of Immunodominant Membrane-Associated 30- to 36-Kilodalton Fraction Antigens of Leishmania infantum Promastigotes, Reacting with Sera from Mediterranean Visceral Leishmaniasis Patients

    PubMed Central

    Kamoun-Essghaier, Sayda; Guizani, Ikram; Strub, Jean Marc; Van Dorsselaer, Alain; Mabrouk, Kamel; Ouelhazi, Lazhar; Dellagi, Koussay

    2005-01-01

    The aim of the present study was to identify and characterize proteins of a 30- to 36-kDa fraction of Leishmania infantum promastigote membranes previously shown to be an immunodominant antigen(s) in Mediterranean visceral leishmaniasis (MVL) and a consistent and reliable serological marker of this disease. By the first approach, Coomassie-stained protein bands (32- and 33-kDa fractions) that specifically reacted by immunoblotting with sera from MVL patients were excised from the gel and submitted to enzymatic digestion to generate peptides. Four peptides were sequenced, three of which were shown to be definitely associated with MVL-reactive antigens and ascribed to a mitochondrial integral ADP-ATP carrier protein from L. major, a putative NADH cytochrome b5 reductase, and a putative mitochondrial carrier protein, respectively. The second approach combined two-dimensional gel electrophoresis of membrane antigens and mass spectrometry (liquid chromatography-mass spectrometry/mass spectrometry) by using a quadrupole time-of-flight analysis. Six immunoreactive spots that resolved within a molecular mass range of 30 to 36 kDa and a pH range of 6.7 to 7.4 corresponded to four Leishmania products. The sequences derived from two spots were ascribed to a beta subunit-like guanine nucleotide binding protein, known as the activated protein kinase C receptor homolog antigen LACK, and to a probable member of the aldehyde reductase family. One spot was identified as a probable ubiquinol-cytochrome c reductase (EC 1.10.2.2) Rieske iron-sulfur protein precursor. The remaining three spots were identified as truncated forms of elongation factor 1α. These antigens correspond to conserved proteins ubiquitously expressed in eukaryotic cells and represent potential candidates for the design of a reliable tool for the diagnosis of this disease. PMID:15699427

  16. Monoclonal antibodies directed against Leishmania secreted acid phosphatase and lipophosphoglycan. Partial characterization of private and public epitopes.

    PubMed

    Ilg, T; Harbecke, D; Wiese, M; Overath, P

    1993-10-15

    Leishmania promastigotes, the stage of the parasite characteristic for the sandfly vector, express an abundant glycoconjugate, called lipophosphoglycan, at their surface. Lipophosphoglycan consists of lysoalkyl-sn-glycerophosphoinositol linked to a phosphosaccharide core conserved in all species, which is connected to PO4-6Gal beta 1,4Man alpha 1 repeats with species-specific substitutions at the Gal residue; the repeats are capped by conserved and species-specific oligosaccharides. Most Leishmania species also secrete an acid phosphatase, which, in Leishmania mexicana, is a filamentous complex composed of a phosphorylated glycoprotein and non-covalently associated proteo-(high-molecular-mass)phosphoglycan. The secreted acid phosphatase complex was used as an antigen to derive a panel of monoclonal antibodies (mAbs). A total of 25 mAbs (17 novel and 8 previously described) were tested by different techniques for their specificity against lipophosphoglycan and secreted acid phosphatase from several Leishmania species. This comparison and the modification of the antigens by chemical or enzymic treatments allowed a classification of the mAbs into several groups. First, from 25 mAbs examined, 22 recognize lipophosphoglycan and the enzyme complex of L. mexicana; only three are specific for secreted acid phosphatase. Two of the latter group are also directed against carbohydrate structures, whereas the third mAb recognizes the 100-kDa polypeptide of the complex. The secreted acid-phosphatase-specific class detects antigen in the flagellar pocket of promastigotes while all anti-lipophosphoglycan mAbs bind to the cell surface. Second, all 15 anti-lipophosphoglycan mAbs investigated in detail appear to be directed against the phosphosaccharide repeats or the cap structure rather than the phosphosaccharide core. Two mAbs recognize terminal cap-structures containing Man alpha 1,2Man residues. Four antibodies are specific for L. mexicana and are probably directed against PO4

  17. An axenic amastigote system for drug screening.

    PubMed Central

    Callahan, H L; Portal, A C; Devereaux, R; Grogl, M

    1997-01-01

    Currently available primary screens for selection of candidate antileishmanial compounds are not ideal. The choices include screens that are designed to closely reflect the situation in vivo but are labor-intensive and expensive (intracellular amastigotes and animal models) and screens that are designed to facilitate rapid testing of a large number of drugs but do not use the clinically relevant parasite stage (promastigote model). The advent of successful in vitro culture of axenic amastigotes permits the development of a primary screen which is quick and easy like the promastigote screen but still representative of the situation in vivo, since it uses the relevant parasite stage. We have established an axenic amastigote drug screening system using a Leishmania mexicana strain (strain M379). A comparison of the 50% inhibitory concentration (IC50) drug sensitivity profiles of M379 promastigotes, intracellular amastigotes, and axenic amastigotes for six clinically relevant antileishmanial drugs (sodium stibogluconate, meglumine antimoniate, pentamidine, paromomycin, amphotericin B, WR6026) showed that M379 axenic amastigotes are a good model for a primary drug screen. Promastigote and intracellular amastigote IC50s differed for four of the six drugs tested by threefold or more; axenic amastigote and intracellular amastigote IC50s differed by twofold for only one drug. This shows that the axenic amastigote susceptibility to clinically used reference drugs is comparable to the susceptibility of amastigotes in macrophages. These data also suggest that for the compounds tested, susceptibility is intrinsic to the parasite stage. This contradicts previous hypotheses that suggested that the activities of antimonial agents against intracellular amastigotes were solely a function of the macrophage. PMID:9087496

  18. Antigenicity, Immunogenicity and Protective Efficacy of Three Proteins Expressed in the Promastigote and Amastigote Stages of Leishmania infantum against Visceral Leishmaniasis

    PubMed Central

    Martins, Vivian Tamietti; Chávez-Fumagalli, Miguel Angel; Lage, Daniela Pagliara; Duarte, Mariana Costa; Garde, Esther; Costa, Lourena Emanuele; da Silva, Viviane Gomes; Oliveira, Jamil Silvano; de Magalhães-Soares, Danielle Ferreira; Teixeira, Santuza Maria Ribeiro; Fernandes, Ana Paula; Soto, Manuel; Tavares, Carlos Alberto Pereira; Coelho, Eduardo Antonio Ferraz

    2015-01-01

    In the present study, two Leishmania infantum hypothetical proteins present in the amastigote stage, LiHyp1 and LiHyp6, were combined with a promastigote protein, IgE-dependent histamine-releasing factor (HRF); to compose a polyproteins vaccine to be evaluated against L. infantum infection. Also, the antigenicity of the three proteins was analyzed, and their use for the serodiagnosis of canine visceral leishmaniasis (CVL) was evaluated. The LiHyp1, LiHyp6, and HRF DNA coding sequences were cloned in prokaryotic expression vectors and the recombinant proteins were purified. When employed in ELISA assays, all proteins were recognized by sera from visceral leishmaniasis (VL) dogs, and presented no cross-reactivity with either sera from dogs vaccinated with a Brazilian commercial vaccine, or sera of Trypanosoma cruzi-infected or Ehrlichia canis-infected animals. In addition, the antigens were not recognized by antibodies from non-infected animals living in endemic or non-endemic areas for leishmaniasis. The immunogenicity and protective efficacy of the three proteins administered in the presence of saponin, individually or in combination (composing a polyproteins vaccine), were evaluated in a VL murine model: BALB/c mice infected with L. infantum. Spleen cells from mice inoculated with the individual proteins or with the polyproteins vaccine plus saponin showed a protein-specific production of IFN-γ, IL-12, and GM-CSF after an in vitro stimulation, which was maintained after infection. These animals presented significant reductions in the parasite burden in different evaluated organs, when compared to mice inoculated with saline or saponin. The decrease in parasite burden was associated with an IL-12-dependent production of IFN-γ against parasite total extracts (produced mainly by CD4+ T cells), correlated to the induction of parasite proteins-driven NO production. Mice inoculated with the recombinant protein-based vaccines showed also high levels of parasite

  19. Antigenicity, Immunogenicity and Protective Efficacy of Three Proteins Expressed in the Promastigote and Amastigote Stages of Leishmania infantum against Visceral Leishmaniasis.

    PubMed

    Martins, Vivian Tamietti; Chávez-Fumagalli, Miguel Angel; Lage, Daniela Pagliara; Duarte, Mariana Costa; Garde, Esther; Costa, Lourena Emanuele; da Silva, Viviane Grazielle; Silva, Viviane Gomes da; Oliveira, Jamil Silvano; Magalhães-Soares, Danielle Ferreira de; Teixeira, Santuza Maria Ribeiro; Fernandes, Ana Paula; Soto, Manuel; Tavares, Carlos Alberto Pereira; Coelho, Eduardo Antonio Ferraz

    2015-01-01

    In the present study, two Leishmania infantum hypothetical proteins present in the amastigote stage, LiHyp1 and LiHyp6, were combined with a promastigote protein, IgE-dependent histamine-releasing factor (HRF); to compose a polyproteins vaccine to be evaluated against L. infantum infection. Also, the antigenicity of the three proteins was analyzed, and their use for the serodiagnosis of canine visceral leishmaniasis (CVL) was evaluated. The LiHyp1, LiHyp6, and HRF DNA coding sequences were cloned in prokaryotic expression vectors and the recombinant proteins were purified. When employed in ELISA assays, all proteins were recognized by sera from visceral leishmaniasis (VL) dogs, and presented no cross-reactivity with either sera from dogs vaccinated with a Brazilian commercial vaccine, or sera of Trypanosoma cruzi-infected or Ehrlichia canis-infected animals. In addition, the antigens were not recognized by antibodies from non-infected animals living in endemic or non-endemic areas for leishmaniasis. The immunogenicity and protective efficacy of the three proteins administered in the presence of saponin, individually or in combination (composing a polyproteins vaccine), were evaluated in a VL murine model: BALB/c mice infected with L. infantum. Spleen cells from mice inoculated with the individual proteins or with the polyproteins vaccine plus saponin showed a protein-specific production of IFN-γ, IL-12, and GM-CSF after an in vitro stimulation, which was maintained after infection. These animals presented significant reductions in the parasite burden in different evaluated organs, when compared to mice inoculated with saline or saponin. The decrease in parasite burden was associated with an IL-12-dependent production of IFN-γ against parasite total extracts (produced mainly by CD4+ T cells), correlated to the induction of parasite proteins-driven NO production. Mice inoculated with the recombinant protein-based vaccines showed also high levels of parasite

  20. Axenically cultured amastigote forms as an in vitro model for investigation of antileishmanial agents.

    PubMed Central

    Sereno, D; Lemesre, J L

    1997-01-01

    Using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide microassay, previously described as a means of quantifying Leishmania amazonensis in vitro at the amastigote stage (D. Sereno and J. L. Lemesre, Parisitol. Res., in press), we have compared the activities of seven drugs, including those currently used to treat leishmaniasis, against axenically grown amastigote and promastigote forms of three Leishmania species (L. amazonensis, L. mexicana, and L. infantum, responsible for diffuse cutaneous, cutaneous, and visceral leishmaniasis, respectively). The ability of axenically cultured amastigote organisms to be used in an investigation of antileishmanial agents was first evaluated. We have confirmed the toxicities of sodium stibogluconate (Pentostam), pentamidine, and amphotericin B to active and dividing populations of axenically cultured amastigotes. The toxicity of potassium antimonyl tartrate trihydrate, which is generally higher than that of Pentostam, seemed to indicate that pentavalent antimony can be metabolized in vivo to compounds, possibly trivalent in nature, which are more active against the amastigote organisms. When the drug susceptibilities of parasites at both stages were compared, great variations were found for all the drugs studied. These major differences, which show the specific chemosusceptibility of the parasite at the mammalian stage, demonstrate the potential of using cultured amastigotes instead of promastigotes in a drug-screening procedure for early detection. This in vitro model may help in the isolation of active compounds, particularly those with low-grade activities, against the mammalian stage of the parasite. PMID:9145854

  1. Identification of a compensatory mutant (lpg2-REV) of Leishmania major able to survive as amastigotes within macrophages without LPG2-dependent glycoconjugates and its significance to virulence and immunization strategies.

    PubMed

    Späth, Gerald F; Lye, Lon-Fye; Segawa, Hiroaki; Turco, Salvatore J; Beverley, Stephen M

    2004-06-01

    Different Leishmania species rely to different extents on abundant glycoconjugates, such as lipophosphoglycan (LPG) and related molecules, in mammalian infections. Previously, we showed that Leishmania major deletion mutants lacking the Golgi GDP-mannose transporter LPG2, which is required for assembly of the dominant phosphoglycan (PG) repeats of LPG, were unable to survive in macrophages. These lpg2- mutants, however, retained the ability to generate asymptomatic, persistent infections in mice. In contrast, Ilg and colleagues showed that Leishmania mexicana LPG2 mutants retained virulence for mice. Here we identified a partial revertant population of the L. major lpg2- mutants (designated lpg2(-)REV) that had regained the ability to replicate in macrophages and induce disease pathology through a compensatory change. Like the lpg2 parent, the lpg2(-)REV revertant was unable to synthesize LPG2-dependent PGs in the promastigote stage and thus remained highly attenuated in the ability to induce infection. However, after considerable delay lpg2(-)REV revertant-infected mice exhibited lesions, and amastigotes isolated from these lesions were able to replicate within macrophages despite the fact that they were unable to synthesize PGs. Thus, in some respects, the lpg2(-)REV amastigotes resemble L. mexicana amastigotes. Future studies of the gene(s) responsible may shed light on the mechanisms employed by L. major to survive in the absence of LPG2-dependent glycoconjugates and may also improve the potential of the lpg2- L. major line to serve as a live parasite vaccine by overcoming its tendency to revert toward virulence. PMID:15155672

  2. Inhibition of trypsin expression in Lutzomyia longipalpis using RNAi enhances the survival of Leishmania

    PubMed Central

    2009-01-01

    Background Leishmania parasites must overcome several barriers to achieve transmission by their sand fly vectors. One of the earliest threats is exposure to enzymes during blood meal digestion. Trypsin-like enzymes appear to be detrimental to parasite survival during the very early phase of development as amastigotes transform into promastigote stages. Here, we investigate whether parasites can affect trypsin secretion by the sand fly midgut epithelium and if inhibition of this process is of survival value to the parasites. Results Infections of Lutzomyia longipalpis with Leishmania mexicana were studied and these showed that infected sand flies produced less trypsin-like enzyme activity during blood meal digestion when compared to uninfected controls. RNA interference was used to inhibit trypsin 1 gene expression by micro-injection into the thorax, as trypsin 1 is the major blood meal induced trypsin activity in the sand fly midgut. Injection of specific double stranded RNA reduced trypsin 1 expression as assessed by RT-PCR and enzyme assays, and also led to increased numbers of parasites in comparison with mock-injected controls. Injection by itself was observed to have an inhibitory effect on the level of infection, possibly through stimulation of a wound repair or immune response by the sand fly. Conclusion Leishmania mexicana was shown to be able to modulate trypsin secretion by Lutzomyia longipalpis to its own advantage, and direct inhibition of trypsin gene expression led to increased parasite numbers in the midguts of infected flies. Successful application of RNA interference methodology to Leishmania-infected sand flies now opens up the use of this technique to study a wide range of sand fly genes and their role in the parasite-vector interaction. PMID:20003192

  3. Sialic acids in different Leishmania sp., its correlation with nitric oxide resistance and host responses.

    PubMed

    Ghoshal, Angana; Gerwig, Gerrit J; Kamerling, Johannis P; Mandal, Chitra

    2010-05-01

    The presence of different derivatives of sialic acids (SA) on Leishmania donovani instigated us to investigate their status on different strains of Leishmania sp. causing different forms of the disease. Leishmania tropica (K27), Leishmania major (JISH118) and Leishmania mexicana (LV4) responsible for cutaneous, Leishmania braziliensis (L280) and Leishmania amazonensis (LV81) causing diffuse and Leishmania infantum (MON29) responsible for visceral leishmaniasis were included in this study. The strains showed a differential distribution of SA in spite of their close resemblance in pathogenesis. K27, JISH118, L280 and MON29 were categorized as high SA-containing strains having enhanced 9-O-acetyl sialic acid (9-O-AcSA(high)) whereas LV4 and LV81 evidenced considerably reduced SA. Interestingly, 9-O-AcSA(high) promastigotes showed significant viability as compared to their de-O-acetylated forms after exposure to NaNO(2) suggesting the involvement of 9-O-AcSA in conferring nitric oxide (NO) resistance. Enhanced intracellular survivability was demonstrated following infection of human macrophages with 9-O-AcSA(high) promastigotes in contrast to their de-O-acetylated forms indicating their contribution in bestowing a survival benefit. Additionally, reduced accumulation of NO, interleukin-12 and interferon-gamma in the supernatant of macrophages infected with 9-O-AcSA(high) promastigotes indicated suppression of leishmanicidal host responses. However, LV4 and LV81 with least 9-O-AcSA, before and after de-O-acetylation, showed unaltered NO resistance, multiplicity and host responses signifying the probable involvement of other determinants which may be a function of their inherent parasitic attribute. Hence, enhanced levels of 9-O-AcSA serve as one of the potential determinants responsible for increased NO resistance and survivability of parasites by inhibition of host responses. PMID:20085901

  4. Host instar susceptibility and selection and interspecific competition of three introduced parasitoids of the mealybug Paracoccus marginatus (Hemiptera: Pseudococccidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Acerophagus papayae, Anagyrus loecki, and Pseudleptomastix mexicana, three introduced parasitoids of Paracoccus marginatus were investigated for host stage susceptibility and sex ratio, host stage suitability, and interspecific competition. All three parasitoid species were able to develop and emer...

  5. Biology, host instar suitability and susceptibility, and interspecific competition of three introduced parasitoids of Paracoccus marginatus (Hemiptera: Pseudococcidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Biology, host stage suitability and susceptibility, and interspecific competition of three previously introduced parasitoids (Acerophagus papayae, Anagyrus loecki, and Pseudleptomastix mexicana) (Hymenoptera: Encyrtidae) of Paracoccus marginatus were studied in the laboratory. Compared to P. mexica...

  6. Developmental time, longevity, and lifetime fertility of three introduced parasitoids of the mealybug Paracoccus marginatus (Hemiptera: Pseudoccidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Developmental time, longevity, and lifetime fertility of three previously introduced parasitoids (Acerophagus papayae Noyes and Schauff, Anagyrus loecki Noyes and Menezes, and Pseudleptomastix mexicana Noyes and Schauff) (Hymenoptera: Encyrtidae) of the mealybug Paracoccus marginatus Williams and Gr...

  7. Diffuse cutaneous leishmaniasis in Mexico.

    PubMed

    Velasco, O; Savarino, S J; Walton, B C; Gam, A A; Neva, F A

    1989-09-01

    In Mexico, 6 cases of diffuse cutaneous leishmaniasis (DCL) were found in widely separated geographic regions. Information was also available on 2 other cases. In addition to the typical clinical features, half of the patients had evidence of nasopharyngeal mucosal involvement. All isolates from the DCL patients were identified as Leishmania mexicana mexicana by isoenzyme analysis and monoclonal antibody typing. In 1 region of Tabasco state where DCL was found, uncomplicated cutaneous leishmaniasis appeared to be highly endemic, and isolates from a few such patients were identified as L. mexicana mexicana. An incidental finding was the recovery of an isolate of L. braziliensis braziliensis from a patient with chiclero ulcer in Oaxaca state. The clinical and epidemiological significance of the reported cases are discussed. PMID:2802018

  8. Th1-stimulatory polyproteins of soluble Leishmania donovani promastigotes ranging from 89.9 to 97.1 kDa offers long-lasting protection against experimental visceral leishmaniasis.

    PubMed

    Kumari, Shraddha; Samant, Mukesh; Misra, Pragya; Khare, Prashant; Sisodia, Brijesh; Shasany, Ajit K; Dube, Anuradha

    2008-10-23

    Our earlier studies identified a fraction (F2) of Leishmania donovani soluble promastigote antigen belonging to 97.4-68 kDa for its ability to stimulate Th1-type cellular responses in cured visceral leishmaniasis (VL) patients as well as in cured hamsters. A further fractionation of F2-fraction into seven subfractions (F2.1-F2.7) and re-assessment for their immunostimulatory responses revealed that out of these, only four (F2.4-F2.7) belonging to 89.9-97.1 kDa, stimulated remarkable Th1-type cellular responses either individually or in a pooled form (P4-7). In this study these potential subfractions were further assessed for their prophylactic potential in combination with BCG against L. donovani challenge in hamsters. Optimum parasite inhibition ( approximately 99%) was obtained in hamsters vaccinated with pooled subfractions and they survived for 1 year. The protection was further supported by remarkable lymphoproliferative, IFN-gamma and IL-12 responses along with profound delayed type hypersensitivity and increased levels of Leishmania-specific IgG2 antibody as observed on days 45, 90 and 120 post-challenge suggesting that a successful subunit vaccine against VL may require multiple Th1-immunostimulatory proteins. MALDI-TOF-MS/MS analysis of these subfractions further revealed that of the 19 identified immunostimulatory proteins, Elongation factor-2, p45, Heat shock protein-70/83, Aldolase, Enolase, Triosephosphate isomerase, Disulfideisomerase and Calreticulin were the major ones in these subfractions. PMID:18762224

  9. "Estudia para que no te pase lo que a mi": Narrativas Culturales Sobre el Valor de la Escuela en Familias Mexicanas ("Study So That What Happened to Me Doesn't Happen to You": Cultural Narratives about the Value of Schooling in Mexican Families).

    ERIC Educational Resources Information Center

    Ramos, Claudia Saucedo

    2003-01-01

    Narratives used by parents in Mexican working-class families to motivate their children to study are examined for the advice offered and parental attitudes about the importance of school. The integration of contemporary pressures about the value of schooling is investigated, as well as the ways in which the value of schooling is being…

  10. Development of a New Antileishmanial Aziridine-2,3-Dicarboxylate-Based Inhibitor with High Selectivity for Parasite Cysteine Proteases.

    PubMed

    Schad, Caroline; Baum, Ulrike; Frank, Benjamin; Dietzel, Uwe; Mattern, Felix; Gomes, Carlos; Ponte-Sucre, Alicia; Moll, Heidrun; Schurigt, Uta; Schirmeister, Tanja

    2016-02-01

    Leishmaniasis is one of the major neglected tropical diseases of the world. Druggable targets are the parasite cysteine proteases (CPs) of clan CA, family C1 (CAC1). In previous studies, we identified two peptidomimetic compounds, the aziridine-2,3-dicarboxylate compounds 13b and 13e, in a series of inhibitors of the cathepsin L (CL) subfamily of the papain clan CAC1. Both displayed antileishmanial activity in vitro while not showing cytotoxicity against host cells. In further investigations, the mode of action was characterized in Leishmania major. It was demonstrated that aziridines 13b and 13e mainly inhibited the parasitic cathepsin B (CB)-like CPC enzyme and, additionally, mammalian CL. Although these compounds induced cell death of Leishmania promastigotes and amastigotes in vitro, the induction of a proleishmanial T helper type 2 (Th2) response caused by host CL inhibition was observed in vivo. Therefore, we describe here the synthesis of a new library of more selective peptidomimetic aziridine-2,3-dicarboxylates discriminating between host and parasite CPs. The new compounds are based on 13b and 13e as lead structures. One of the most promising compounds of this series is compound s9, showing selective inhibition of the parasite CPs LmaCatB (a CB-like enzyme of L. major; also named L. major CPC) and LmCPB2.8 (a CL-like enzyme of Leishmania mexicana) while not affecting mammalian CL and CB. It displayed excellent leishmanicidal activities against L. major promastigotes (50% inhibitory concentration [IC50] = 37.4 μM) and amastigotes (IC50 = 2.3 μM). In summary, we demonstrate a new selective aziridine-2,3-dicarboxylate, compound s9, which might be a good candidate for future in vivo studies. PMID:26596939

  11. Development of a New Antileishmanial Aziridine-2,3-Dicarboxylate-Based Inhibitor with High Selectivity for Parasite Cysteine Proteases

    PubMed Central

    Schad, Caroline; Baum, Ulrike; Frank, Benjamin; Dietzel, Uwe; Mattern, Felix; Gomes, Carlos; Ponte-Sucre, Alicia; Moll, Heidrun

    2015-01-01

    Leishmaniasis is one of the major neglected tropical diseases of the world. Druggable targets are the parasite cysteine proteases (CPs) of clan CA, family C1 (CAC1). In previous studies, we identified two peptidomimetic compounds, the aziridine-2,3-dicarboxylate compounds 13b and 13e, in a series of inhibitors of the cathepsin L (CL) subfamily of the papain clan CAC1. Both displayed antileishmanial activity in vitro while not showing cytotoxicity against host cells. In further investigations, the mode of action was characterized in Leishmania major. It was demonstrated that aziridines 13b and 13e mainly inhibited the parasitic cathepsin B (CB)-like CPC enzyme and, additionally, mammalian CL. Although these compounds induced cell death of Leishmania promastigotes and amastigotes in vitro, the induction of a proleishmanial T helper type 2 (Th2) response caused by host CL inhibition was observed in vivo. Therefore, we describe here the synthesis of a new library of more selective peptidomimetic aziridine-2,3-dicarboxylates discriminating between host and parasite CPs. The new compounds are based on 13b and 13e as lead structures. One of the most promising compounds of this series is compound s9, showing selective inhibition of the parasite CPs LmaCatB (a CB-like enzyme of L. major; also named L. major CPC) and LmCPB2.8 (a CL-like enzyme of Leishmania mexicana) while not affecting mammalian CL and CB. It displayed excellent leishmanicidal activities against L. major promastigotes (50% inhibitory concentration [IC50] = 37.4 μM) and amastigotes (IC50 = 2.3 μM). In summary, we demonstrate a new selective aziridine-2,3-dicarboxylate, compound s9, which might be a good candidate for future in vivo studies. PMID:26596939

  12. Modulation of Leishmania major aquaglyceroporin activity by a mitogen-activated protein kinase

    PubMed Central

    Mandal, Goutam; Sharma, Mansi; Kruse, Martin; Sander-Juelch, Claudia; Munro, Laura Anne; Wang, Yong; Vilg, Jenny Veide; Tamás, Markus J; Bhattacharjee, Hiranmoy; Wiese, Martin; Mukhopadhyay, Rita

    2012-01-01

    Summary Leishmania major aquaglyceroporin (LmjAQP1) adventitiously facilitates the uptake of antimonite [Sb(III)], an active form of Pentostam® or Glucantime®, which are the first line of defense against all forms of leishmaniasis. The present paper shows that LmjAQP1 activity is modulated by the mitogen-activated protein kinase, LmjMPK2. Leishmania parasites co-expressing LmjAQP1 and LmjMPK2 show increased Sb(III) uptake and increased Sb(III) sensitivity. When subjected to a hypo-osmotic stress, these cells show faster volume recovery than cells expressing LmjAQP1 alone. LmjAQP1 is phosphorylated in vivo at Thr197 and this phosphorylation requires LmjMPK2 activity. Lys42 of LmjMPK2 is critical for its kinase activity. Cells expressing altered T197A LmjAQP1 or K42A LmjMPK2 showed decreased Sb(III) influx and a slower volume recovery than cells expressing wild type proteins. Phosphorylation of LmjAQP1 led to a decrease in its turnover rate affecting LmjAQP1 activity. Although LmjAQP1 is localized to the flagellum of promastigotes, upon phosphorylation, it is relocalized to the entire surface of the parasite. L. mexicana promastigotes with an MPK2 deletion showed reduced Sb(III) uptake and slower volume recovery than wild type cells. This is the first report where a parasite aquaglyceroporin activity is post-translationally modulated by a MAP kinase. PMID:22779703

  13. Bioactive indole alkaloids isolated from Alstonia angustifolia

    PubMed Central

    Pan, Li; Terrazas, César; Muñoz Acuña, Ulyana; Ninh, Tran Ngoc; Chai, Heebyung; Carcache de Blanco, Esperanza J.; Soejarto, Djaja D.; Satoskar, Abhay R.

    2014-01-01

    Bioassay-guided fractionation was conducted on a CHCl3-soluble extract of the stem bark of Alstonia angustifolia (Apocynaceae) collected in Vietnam using the HT-29 human colon cancer cell line, and led to the isolation of a new sarpagine-type indole alkaloid (1), together with nine known alkaloids, including four macroline-derived alkaloids (2–5), a sarpagine-type alkaloid (6), and four macroline-pleiocarpamine bisindole alkaloids (7–10). The structure of the new compound (1) was determined on the basis of spectroscopic data interpretation. Compounds 1–10 were evaluated in vitro for their NF-κB (p65) inhibitory activity against the Hela cells in an ELISA assay. The new sarpagine alkaloid, N(4)-methyltalpinine (1), was found to show significant NF-κB inhibitory activity (ED50 = 1.2 µM). Furthermore, all the isolates (1–10) were evaluated in vitro for their antileishmanial activity, and compounds (1–4, 6 and 8–10) exhibited leishmaniacidal activity against promastigotes of Leishmania mexicana. PMID:25584095

  14. A putative novel nuclear-encoded subunit of the cytochrome c oxidase complex in trypanosomatids.

    PubMed

    Maslov, Dmitri A; Zíková, Alena; Kyselová, Iveta; Lukes, Julius

    2002-01-01

    A relatively large nuclear-encoded polypeptide, designated trCOIV, is found in the cytochrome c oxidase (CO) complex of trypanosomatids. In order to determine if this polypeptide represents a bona fide subunit of the complex, we have characterized the cDNA and the gene for this polypeptide in Leishmania tarentolae. Its nuclear gene has no sequence similarity to mammalian COIV. The trCOIV preprotein has a long mitochondrial targeting sequence of 31 residues. The mature polypeptide cofractionates with kinetoplast-mitochondria and its preferential mitochondrial localization was confirmed by immunofluorescence and immunoelectron microscopy. Based on the hydropathy plot analysis, the protein lacks pronounced transmembrane domains and likely occupies a peripheral position within the CO complex. The corresponding genes are also present in the sequenced portions of the Trypanosoma cruzi, Trypanosoma brucei and Leishmania major genomes, and the same polypeptide is found in cytochrome oxidase isolated from procyclic T. brucei and promastigote Leishmania mexicana amazonensis. However, the trCOIV gene, the mRNA and the polypeptide could not be detected in a respiration-deficient trypanosomatid Phytomonas serpens. PMID:12467979

  15. Metabolomic Analyses of Leishmania Reveal Multiple Species Differences and Large Differences in Amino Acid Metabolism

    PubMed Central

    Wang, Lijie; Zhang, Tong; Watson, David G.; Silva, Ana Marta; Coombs, Graham H.

    2015-01-01

    Comparative genomic analyses of Leishmania species have revealed relatively minor heterogeneity amongst recognised housekeeping genes and yet the species cause distinct infections and pathogenesis in their mammalian hosts. To gain greater information on the biochemical variation between species, and insights into possible metabolic mechanisms underpinning visceral and cutaneous leishmaniasis, we have undertaken in this study a comparative analysis of the metabolomes of promastigotes of L. donovani, L. major and L. mexicana. The analysis revealed 64 metabolites with confirmed identity differing 3-fold or more between the cell extracts of species, with 161 putatively identified metabolites differing similarly. Analysis of the media from cultures revealed an at least 3-fold difference in use or excretion of 43 metabolites of confirmed identity and 87 putatively identified metabolites that differed to a similar extent. Strikingly large differences were detected in their extent of amino acid use and metabolism, especially for tryptophan, aspartate, arginine and proline. Major pathways of tryptophan and arginine catabolism were shown to be to indole-3-lactate and arginic acid, respectively, which were excreted. The data presented provide clear evidence on the value of global metabolomic analyses in detecting species-specific metabolic features, thus application of this technology should be a major contributor to gaining greater understanding of how pathogens are adapted to infecting their hosts. PMID:26368322

  16. Comparative effects of 4-aminopyrazolopyrimidine, its 2'-deoxyriboside derivative, and allopurinol on in vitro growth of American Leishmania species.

    PubMed Central

    Avila, J L; Casanova, M A

    1982-01-01

    When assayed on promastigotes of 10 American Leishmania isolates (including Leishmania brasiliensis and mexicana species), 4-aminopyrazolopyrimidine (AAP) was severalfold more active than allopurinol as a leishmanistatic drug. There were some intraspecific and interspecific differences among the isolates in their susceptibility to the inhibitory effects of APP and allopurinol. APP-2'-deoxyriboside did not affect the 10 isolates tested. This was surprising, because allopurinol riboside has previously been shown to be more active than allopurinol. In all of the American Leishmania isolates tested, the metabolism of [14C]6-APP resulted in a high level of HPP-ribose-5'-P and lower levels of APP-ribose-5'-P, APP-ribose-5'-PP, and APP-ribose-5'-PPP. MP, LR, LBY, and JAP isolates strongly converted APP into APP derivatives, thus perhaps explaining their greater susceptibility to the inhibitory effects of APP. With the 10 American Leishmania isolates tested, several purines reversed the inhibitory effects of allopurinol, but only adenine countered the inhibitory effects of APP. This suggests biochemical differences in the mechanisms of action of allopurinol and APP. Finally, and contrary to previous claims, the reversal by purines of the allopurinol-induced growth inhibition was not a Leishmania species-specific effect. PMID:6982678

  17. High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species

    PubMed Central

    Zampieri, Ricardo Andrade; Laranjeira-Silva, Maria Fernanda; Muxel, Sandra Marcia; Stocco de Lima, Ana Carolina; Shaw, Jeffrey Jon; Floeter-Winter, Lucile Maria

    2016-01-01

    Background Protozoan parasites of the genus Leishmania cause a large spectrum of clinical manifestations known as Leishmaniases. These diseases are increasingly important public health problems in many countries both within and outside endemic regions. Thus, an accurate differential diagnosis is extremely relevant for understanding epidemiological profiles and for the administration of the best therapeutic protocol. Methods/Principal Findings Exploring the High Resolution Melting (HRM) dissociation profiles of two amplicons using real time polymerase chain reaction (real-time PCR) targeting heat-shock protein 70 coding gene (hsp70) revealed differences that allowed the discrimination of genomic DNA samples of eight Leishmania species found in the Americas, including Leishmania (Leishmania) infantum chagasi, L. (L.) amazonensis, L. (L.) mexicana, L. (Viannia) lainsoni, L. (V.) braziliensis, L. (V.) guyanensis, L. (V.) naiffi and L. (V.) shawi, and three species found in Eurasia and Africa, including L. (L.) tropica, L. (L.) donovani and L. (L.) major. In addition, we tested DNA samples obtained from standard promastigote culture, naturally infected phlebotomines, experimentally infected mice and clinical human samples to validate the proposed protocol. Conclusions/Significance HRM analysis of hsp70 amplicons is a fast and robust strategy that allowed for the detection and discrimination of all Leishmania species responsible for the Leishmaniases in Brazil and Eurasia/Africa with high sensitivity and accuracy. This method could detect less than one parasite per reaction, even in the presence of host DNA. PMID:26928050

  18. Leishmania amazonensis Arginase Compartmentalization in the Glycosome Is Important for Parasite Infectivity

    PubMed Central

    da Silva, Maria Fernanda Laranjeira; Zampieri, Ricardo Andrade; Muxel, Sandra M.; Beverley, Stephen M.; Floeter-Winter, Lucile M.

    2012-01-01

    In Leishmania, de novo polyamine synthesis is initiated by the cleavage of L-arginine to urea and L-ornithine by the action of arginase (ARG, E.C. 3.5.3.1). Previous studies in L. major and L. mexicana showed that ARG is essential for in vitro growth in the absence of polyamines and needed for full infectivity in animal infections. The ARG protein is normally found within the parasite glycosome, and here we examined whether this localization is required for survival and infectivity. First, the localization of L. amazonensis ARG in the glycosome was confirmed in both the promastigote and amastigote stages. As in other species, arg− L. amazonensis required putrescine for growth and presented an attenuated infectivity. Restoration of a wild type ARG to the arg− mutant restored ARG expression, growth and infectivity. In contrast, restoration of a cytosol-targeted ARG lacking the glycosomal SKL targeting sequence (argΔSKL) restored growth but failed to restore infectivity. Further study showed that the ARGΔSKL protein was found in the cytosol as expected, but at very low levels. Our results indicate that the proper compartmentalization of L. amazonensis arginase in the glycosome is important for enzyme activity and optimal infectivity. Our conjecture is that parasite arginase participates in a complex equilibrium that defines the fate of L-arginine and that its proper subcellular location may be essential for this physiological orchestration. PMID:22479507

  19. Complement binding to Leishmania donovani promastigotes (LD)

    SciTech Connect

    Puentes, S.M.; Bates, P.A.; Dwyer, D.M.; Joiner, K.A.

    1986-03-01

    To study the binding and processing of C3 on LD, parasites in various phases of growth were incubated in human serum deficient in complement component 8 containing /sup 125/I-C3. Uptake of /sup 125/I-C3 is rapid, peaking at 1.7-2.1 x 10/sup 6/ C3 molecules bound per parasite at 15 minutes for all growth phases, and decreases thereafter with continued incubation. One half of total C3 bound is spontaneously released by 90 minutes of incubation with all LD phases and occurs at a similar rate for LD washed free of serum and incubated at 37/sup 0/ C in buffer. As assessed by SDS-PAGE autoradiography, C3 on the surface of LD is present as C3b (36 to 50%) and iC3b (50 to 65%), linked covalently via a bond resistant to hydroxylamine treatment, presumably an amide linkage. Immunoblot analysis of purified membranes from serum-incubated LD, using rabbit antibody to C3 and LD surface constituents, strongly suggests that a major C3 acceptor is the LD acid phosphatase (AP). These results, in conjunction with recent studies, suggest a previously unrecognized role of AP as a C3 acceptor and, thus, as a molecule potentially involved in parasite binding and uptake.

  20. In vitro life cycle of pentamidine-resistant amastigotes: stability of the chemoresistant phenotypes is dependent on the level of resistance induced.

    PubMed Central

    Sereno, D; Lemesre, J L

    1997-01-01

    Using a continuous drug pressure protocol, we induced pentamidine resistance in an active and dividing population of amastigote forms of Leishmania mexicana. We selected in vitro two clones with different levels of resistance to pentamidine, with clone LmPENT5 being resistant to 5 microM pentamidine, while clone LmPENT20 was resistant to 20 microM pentamidine. Resistance indexes (50% inhibitory concentration [IC50] after drug presure/IC50 before drug pressure) of 2 (LmPENT5) and 6 (LmPENT20) were determined after drug selection. Both resistant clones expressed significant cross-resistance to diminazene aceturate and primaquine. Pentamidine resistance was not reversed by verapamil, a calcium channel blocker known to reverse multidrug resistance (A. J. Bitonti, et al., Science 242:1301-1303, 1988; A. R. C. Safa et al., J. Biol. Chem. 262:7884-7888, 1987). No difference in the in vitro infectivity for resident mouse macrophages was observed between the wild-type clone (clone LmWT) and pentamidine-resistant clones. During in vitro infectivity experiments, when the life cycle was performed starting from the intramacrophagic amastigote stage, the drug resistance of the resulting LmPENT20 amastigotes was preserved even if the intermediate promastigote stage could not be considered resistant to 20 microM pentamidine. In the same way, when a complete developmental sequence of L. mexicana was achieved axenically by manipulation of appropriate culture conditions, the resulting axenically grown LmPENT20 amastigotes remained pentamidine resistant, whereas LmPENT5 amastigotes lost their ability to resist pentamidine, with IC50s and index of resistance values close to those for the LmWT clone. These results strongly indicate that the level of pentamidine tolerated by resistant amastigotes after the life cycle was dependent on the induced level of resistance. This fact could be significant in the in vivo transmission of drug-resistant parasites by Phlebotominae. Particular

  1. The structure of Leishmania major amastigote lipophosphoglycan.

    PubMed

    Moody, S F; Handman, E; McConville, M J; Bacic, A

    1993-09-01

    Intracellular amastigotes of Leishmania major produce 6 x 10(4) copies/cell of a lipophosphoglycan (LPG) that is structurally distinct from the LPG produced by the extracellular promastigote form of L. major, Leishmania donovani, and Leishmania mexicana (reviewed by McConville, M. J. (1991) Cell Biol. Int. Rep. 15, 779-798). L. major amastigote LPG is composed of a lysoalkyl phosphatidylinositol lipid anchor that links via a diphosphorylated hexasaccharide core to a phosphoglycan (6-100 kDa). The structures of the anchor, the core, and the phosphoglycan were determined by monosaccharide and linkage analysis, fast atom bombardment-mass spectrometry, one-dimensional 1H NMR spectroscopy, and exoglycosidase microsequencing. The lipid anchor contains predominantly 1-O-alkylglycerols with 24:0 and 22:0 alkyl chains. The lipids are linked via a glycerol-myo-inositol-PO4 to a core glycan with the structure -PO4-6)Gal(alpha 1-)Gal(alpha 1-) Galf(beta 1-)[Glc(alpha 1-PO4-)]Man(alpha 1-)Man(alpha 1-)GlcN(alpha 1-). The chromatographic characteristics of the core glycan suggest that the saccharide components are linked similarly in amastigote and promastigote LPG. The phosphoglycan attached to the core consists of -PO4-6)Gal(beta 1-4)Man(alpha 1- repeats units which are either unsubstituted (70%) or substituted (30%) at the 3-position of the Gal residues with oligosaccharide side chains containing primarily Gal and some Glc. Thirteen different types of side chains were identified with the structures [Gal(beta 1-3)]x, where x = 1-11, or Glc(1-3)Glc(1-3), or Glc(1-3)Gal(beta 1-3), where glucose is probably in the beta-configuration. All monosaccharides in the phosphoglycan domain are in the pyranose configuration. The average number of repeat units per molecule is 36. The nonreducing terminus of the phosphoglycan chains probably terminates predominantly in the neutral disaccharide Gal(beta 1-4)Man(alpha 1-. Comparison of the structure of L. major amastigote LPG to L. major

  2. The Comparative Genomics and Phylogenomics of Leishmania amazonensis Parasite

    PubMed Central

    Tschoeke, Diogo A; Nunes, Gisele L; Jardim, Rodrigo; Lima, Joana; Dumaresq, Aline SR; Gomes, Monete R; de Mattos Pereira, Leandro; Loureiro, Daniel R; Stoco, Patricia H; de Matos Guedes, Herbert Leonel; de Miranda, Antonio Basilio; Ruiz, Jeronimo; Pitaluga, André; Silva, Floriano P; Probst, Christian M; Dickens, Nicholas J; Mottram, Jeremy C; Grisard, Edmundo C; Dávila, Alberto MR

    2014-01-01

    Leishmaniasis is an infectious disease caused by Leishmania species. Leishmania amazonensis is a New World Leishmania species belonging to the Mexicana complex, which is able to cause all types of leishmaniasis infections. The L. amazonensis reference strain MHOM/BR/1973/M2269 was sequenced identifying 8,802 codifying sequences (CDS), most of them of hypothetical function. Comparative analysis using six Leishmania species showed a core set of 7,016 orthologs. L. amazonensis and Leishmania mexicana share the largest number of distinct orthologs, while Leishmania braziliensis presented the largest number of inparalogs. Additionally, phylogenomic analysis confirmed the taxonomic position for L. amazonensis within the “Mexicana complex”, reinforcing understanding of the split of New and Old World Leishmania. Potential non-homologous isofunctional enzymes (NISE) were identified between L. amazonensis and Homo sapiens that could provide new drug targets for development. PMID:25336895

  3. Trypanosoma brucei aquaglyceroporin 2 is a high-affinity transporter for pentamidine and melaminophenyl arsenic drugs and the main genetic determinant of resistance to these drugs

    PubMed Central

    Munday, Jane C.; Eze, Anthonius A.; Baker, Nicola; Glover, Lucy; Clucas, Caroline; Aguinaga Andrés, David; Natto, Manal J.; Teka, Ibrahim A.; McDonald, Jennifer; Lee, Rebecca S.; Graf, Fabrice E.; Ludin, Philipp; Burchmore, Richard J. S.; Turner, C. Michael R.; Tait, Andy; MacLeod, Annette; Mäser, Pascal; Barrett, Michael P.; Horn, David; De Koning, Harry P.

    2014-01-01

    Objectives Trypanosoma brucei drug transporters include the TbAT1/P2 aminopurine transporter and the high-affinity pentamidine transporter (HAPT1), but the genetic identity of HAPT1 is unknown. We recently reported that loss of T. brucei aquaglyceroporin 2 (TbAQP2) caused melarsoprol/pentamidine cross-resistance (MPXR) in these parasites and the current study aims to delineate the mechanism by which this occurs. Methods The TbAQP2 loci of isogenic pairs of drug-susceptible and MPXR strains of T. brucei subspecies were sequenced. Drug susceptibility profiles of trypanosome strains were correlated with expression of mutated TbAQP2 alleles. Pentamidine transport was studied in T. brucei subspecies expressing TbAQP2 variants. Results All MPXR strains examined contained TbAQP2 deletions or rearrangements, regardless of whether the strains were originally adapted in vitro or in vivo to arsenicals or to pentamidine. The MPXR strains and AQP2 knockout strains had lost HAPT1 activity. Reintroduction of TbAQP2 in MPXR trypanosomes restored susceptibility to the drugs and reinstated HAPT1 activity, but did not change the activity of TbAT1/P2. Expression of TbAQP2 sensitized Leishmania mexicana promastigotes 40-fold to pentamidine and >1000-fold to melaminophenyl arsenicals and induced a high-affinity pentamidine transport activity indistinguishable from HAPT1 by Km and inhibitor profile. Grafting the TbAQP2 selectivity filter amino acid residues onto a chimeric allele of AQP2 and AQP3 partly restored susceptibility to pentamidine and an arsenical. Conclusions TbAQP2 mediates high-affinity uptake of pentamidine and melaminophenyl arsenicals in trypanosomes and TbAQP2 encodes the previously reported HAPT1 activity. This finding establishes TbAQP2 as an important drug transporter. PMID:24235095

  4. In Vitro and In Vivo Efficacy of Ether Lipid Edelfosine against Leishmania spp. and SbV-Resistant Parasites

    PubMed Central

    Yepes, Edward; Müller, Ingrid; Modolell, Manuel; Muñoz, Diana L.; Robledo, Sara M.; Muskus, Carlos E.; López-Abán, Julio; Muro, Antonio; Vélez, Iván D.; Mollinedo, Faustino

    2012-01-01

    Background The leishmaniases are a complex of neglected tropical diseases caused by more than 20 Leishmania parasite species, for which available therapeutic arsenal is scarce and unsatisfactory. Pentavalent antimonials (SbV) are currently the first-line pharmacologic therapy for leishmaniasis worldwide, but resistance to these compounds is increasingly reported. Alkyl-lysophospoholipid analogs (ALPs) constitute a family of compounds with antileishmanial activity, and one of its members, miltefosine, has been approved as the first oral treatment for visceral and cutaneous leishmaniasis. However, its clinical use can be challenged by less impressive efficiency in patients infected with some Leishmania species, including L. braziliensis and L. mexicana, and by proneness to develop drug resistance in vitro. Methodology/Principal Findings We found that ALPs ranked edelfosine>perifosine>miltefosine>erucylphosphocholine for their antileishmanial activity and capacity to promote apoptosis-like parasitic cell death in promastigote and amastigote forms of distinct Leishmania spp., as assessed by proliferation and flow cytometry assays. Effective antileishmanial ALP concentrations were dependent on both the parasite species and their development stage. Edelfosine accumulated in and killed intracellular Leishmania parasites within macrophages. In vivo antileishmanial activity was demonstrated following oral treatment with edelfosine of mice and hamsters infected with L. major, L. panamensis or L. braziliensis, without any significant side-effect. Edelfosine also killed SbV-resistant Leishmania parasites in in vitro and in vivo assays, and required longer incubation times than miltefosine to generate drug resistance. Conclusions/Significance Our data reveal that edelfosine is the most potent ALP in killing different Leishmania spp., and it is less prone to lead to drug resistance development than miltefosine. Edelfosine is effective in killing Leishmania in culture and within

  5. Immunopathology of experimental cutaneous leishmaniasis.

    PubMed Central

    Andrade, Z. A.; Reed, S. G.; Roters, S. B.; Sadigursky, M.

    1984-01-01

    Relatively susceptible BALB/c and relatively resistant A/J mice were infected subcutaneously in the right hind footpad with promastigotes of Leishmania mexicana amazonensis. A large localized lesion developed within 2 months after infection in the BALB/c mice, while A/J mice exhibited a small discrete fibrotic nodule. Sequential immunologic and histologic examination demonstrated that BALB/c mice developed a nodular foam-cell type of lesion and progressive depression of a delayed-type hypersensitivity (DTH) response to leishmania antigen, while the A/J mice had a mixed cellular fibrosing and encapsulating reaction and developed and maintained positive DTH responses to leishmania antigen. Anti-leishmania antibody responses were positive at similar levels in both strains. The lesions in BALB/c mice were found in bone marrow, tendon, skin appendages, and regional lymph nodes, with a tendency toward cutaneous metastases. Lesions in A/J mice remained localized. Fibrosis, focal fibrinoid necrosis, and lymphocytic and macrophagic infiltration were the outstanding features. Light and transmission electron microscopic studies indicated that no outstanding destruction of leishmanias seemed to occur within macrophages of either mouse strain. In the more resistant A/J mice, however, parasitized macrophages were frequently necrotic, and degenerating leishmanias were often seen free in the interstitial tissue. These observations help the interpretation of the histologic features, as well as the pathogenesis, of cutaneous and mucocutaneous leishmaniasis in man. Images Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 PMID:6691411

  6. Field assessment of three introduced parasitoids of Paracoccus marginatus (Hemiptera: Pseudococcidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Abstract: Field assessment of the ability of three introduced parasitoids (Acerophagus papayae Noyes and Schauff, Anagyrus loecki Noyes, and Pseudleptomastix mexicana Noyes and Schauff) to control Paracoccus marginatus Williams and Granara de Willink was investigated in 2005 and 2006 in three locat...

  7. Efficiency and establishment of three introduced parasitoids of the mealybug Paracoccus marginatus (Hemiptera: Pseudococcidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A study on the efficiency and establishment of three previously introduced parasitoids (Acerophagus papayae, Anagyrus loecki, and Pseudleptomastix mexicana) to control the mealybug Paracoccus marginatus was made in 2005 and 2006, at three locations in Homestead (Miami-Dade County), Florida. In each ...

  8. Preface to "Should animal welfare be law or market driven?"

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Bioethics Symposium, entitled “Should animal welfare be law or market driven?” was held at the joint annual meeting of the American Dairy Science Association, American Society of Animal Science, Poultry Science Association, Asociación Mexicana de Producción Animal, and Canadian Society of Animal...

  9. "Mexico in Transition." Curriculum Units.

    ERIC Educational Resources Information Center

    Oregon Univ., Eugene. Foreign Language Resource Center.

    These curriculum units were developed in a National Endowment for the Humanities 1994 summer seminar "Mexico in Transition." The 23 lessons are written in Spanish. Lessons are entitled: (1) "La Migracion Mexicana Vista a Traves del Cuento 'Paso del Norte' de Juan Rulfo" (Jose Jorge Armendariz); (2) "Los Grupos Indigenas de Mexico: Condiciones…

  10. Leluthia astigma (Ashmead)(Hymenoptera: Braconidae: Doryctinae)as a parasitoid of Agrilus planipennis Fairmaire(Coleoptera: Buprestidae: Agrilinae),with assessment of host associations for Nearctic species of Leluthia Cameron

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The validity of host and host plant records are assessed for Leluthia astigma (Ashmead), Leluthia floridensis Marsh, and Leluthia mexicana Cameron, the three known species of Leluthia Cameron in the Nearctic Region. Leluthia astigma is reported from the exotic invasive pest Agrilus planipennis Fairm...

  11. Mexico: Neighbor in Transition. Foreign Policy Association Headline Series, No. 267.

    ERIC Educational Resources Information Center

    Smith, Peter H.

    One of a series of booklets on world issues, this five-part document presents information about Mexico. Part one examines Mexican history from Cortez to Madero. Emphasis is placed on the revolution era (1910-1920) and the creation of two political parties: the Partido de la Revolucion Mexicana (Mexican Revolutionary Party or PRM) and the Partido…

  12. Triennial Growth Symposium: Dietary regulation of growth development

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The 2010 Triennial Growth Symposium was held immediately before the Joint Annual Meeting of the American Dairy Science Association, Poultry Science Association, Asociación Mexicana de Producción Animal, Canadian Society of Animal Science, Western Section American Society of Animal Science, and Ameri...

  13. The Geography Olympiad in Mexico

    ERIC Educational Resources Information Center

    Garcia-Garcia, Fernando

    2007-01-01

    The formal organisation of science Olympiads in Mexico dates from 1987, when a national contest on Mathematics was held in order to identify a team to represent the country in the International Mathematics Olympiad. In 1991, the Mexican Academy of Sciences ("Academia Mexicana de Ciencias"-AMC) created the National Science Olympiads Program with…

  14. Sheep symposium: Biology and management of low-input lambing in easy-care systems

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Low-input lambing management was the focus of the 2007 Sheep Symposium at the joint annual meetings of the American Society of Animal Science, the American Dairy Science Association, the Asociacio´n Mexicana de Produccio´n Animal, and the Poultry Science Association held in San Antonio, Texas, on Ju...

  15. Identification of potential plant extracts for anti-tick activity against acaricide resistant cattle ticks, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Ghosh, Srikanta; Tiwari, Shashi Shankar; Kumar, Bhanu; Srivastava, Sharad; Sharma, Anil Kumar; Kumar, Sachin; Bandyopadhyay, A; Julliet, Sanis; Kumar, Rajesh; Rawat, A K S

    2015-05-01

    To develop an eco-friendly tick control method, seven plant extracts were prepared using 50 and 95% ethanol and evaluated for acaricidal activity against cattle tick, Rhipicephalus (Boophilus) microplus. The adult immersion test was adopted for testing different extracts. Based on 72 h screening criterion, 95% ethanolic extracts of Datura metel fruits and Argemone mexicana whole plant were found effective showing more than 50% mortality of treated ticks. The 95% ethanolic extracts of D. metel fruits and A. mexicana whole plant exhibited acaricidal and reproductive inhibitory effects on treated ticks. The LC90 values of D. metel and A. mexicana extracts were determined as 7.13 and 11.3%, respectively. However, although both the extracts were found efficacious against deltamethrin-resistant IVRI-4 and multi-acaricide resistant IVRI-5 lines of R. (B.) microplus, they caused less mortality than treated ticks of the reference IVRI-I line. Phytochemical studies indicated the presence of alkaloids and glucosides in D. metel fruits and alkaloids, terpenoids, flavonoids and phenolics in A. mexicana whole plant extracts. The results indicated that these botanicals may play an important role in reducing the use of chemicals for tick control and possibly to manage resistant tick population in environment friendly manner. PMID:25717008

  16. Ionized Gaseous Nebulae

    NASA Astrophysics Data System (ADS)

    Franco, José; Henney, William; Martos, Marco; Peña, Miriam

    2001-06-01

    Conference was held in Mexico City on 2000 November 21-24. Proceedings will be edited by José Franco, William Henney, Marco Martos, and Miriam Peña and will be published in Revista Mexicana de Astronomía y Astrofísica (Serie de Conferencias).

  17. Agricultural Policy and Child Health in Rural Mexico.

    ERIC Educational Resources Information Center

    Brown, Antoinette B.; Partridge, William L.

    1987-01-01

    Reports on the impact on malnutrition of Mexico's Sistema de Alementacion Mexicana (SAM) program, which tried to make the country self-sufficient in basic foods. Concludes that the food problem in Mexico is not so much one of production as it is a problem of distribution and consumption. (PS)

  18. Development in Mexico and Central America. Fulbright-Hays Summer Seminars Abroad Program. Summer 1991.

    ERIC Educational Resources Information Center

    Institute of International Education, New York, NY.

    This document features writings and curriculum projects by teachers who traveled to Mexico and Central America in the summer of 1991 as members of a Fulbright-Hays Summer Seminar. The following items are among the 20 included: Curriculum Project: "'Escritoras Mexicanas Contemporaneas': A Survey of Mexican Women Fiction Writers" (Laura J. Beard);…

  19. Genetic Diversity and Population Structure of Teosinte

    PubMed Central

    Fukunaga, Kenji; Hill, Jason; Vigouroux, Yves; Matsuoka, Yoshihiro; Sanchez G., Jesus; Liu, Kejun; Buckler, Edward S.; Doebley, John

    2005-01-01

    The teosintes, the closest wild relatives of maize, are important resources for the study of maize genetics and evolution and for plant breeding. We genotyped 237 individual teosinte plants for 93 microsatellites. Phylogenetic relationships among species and subspecific taxa were largely consistent with prior analyses for other types of molecular markers. Plants of all species formed monophyletic clades, although relationships among species were not fully resolved. Phylogenetic analysis indicated that the Mexican annual teosintes divide into two clusters that largely correspond to the previously defined subspecies, Z. mays ssp. parviglumis and ssp. mexicana, although there are a few samples that represent either evolutionary intermediates or hybrids between these two subspecies. The Mexican annual teosintes show genetic substructuring along geographic lines. Hybridization or introgression between some teosintes and maize occurs at a low level and appears most common with Z. mays ssp. mexicana. Phylogeographic and phylogenetic analyses of the Mexican annual teosintes indicated that ssp. parviglumis diversified in the eastern part of its distribution and spread from east to west and that ssp. mexicana diversified in the Central Plateau of Mexico and spread along multiple paths to the north and east. We defined core sets of collections of Z. mays ssp. mexicana and ssp. parviglumis that attempt to capture the maximum number of microsatellite alleles for given sample sizes. PMID:15687282

  20. Trinational Library Forum [Proceedings] = Memorias del Foro Trinacional de Bibliotecas = [Actes de] Forum Trinational Des Bibliotheques (4th, Monterrey, Mexico, February 24-26, 1994).

    ERIC Educational Resources Information Center

    Instituto Tecnologico de Monterrey (Mexico).

    At the fourth Trinational Library Forum in 1994, librarians from the United States, Canada, and Mexico gathered to discuss topics of international librarianship. Presentations, some in English and some in Spanish with a brief English language summary, include: (1) "Perspectivas de la Produccion y Exportaciones de la Industria Editorial Mexicana"…

  1. Plastid DNA sequencing and nuclear SNP genotyping help resolve the puzzle of central American Platanus

    PubMed Central

    De Castro, Olga; Di Maio, Antonietta; Lozada García, José Armando; Piacenti, Danilo; Vázquez-Torres, Mario; De Luca, Paolo

    2013-01-01

    Background and Aims Recent research on the history of Platanus reveals that hybridization phenomena occurred in the central American species. This study has two goals: to help resolve the evolutive puzzle of central American Platanus, and to test the potential of real-time polymerase chain reaction (PCR) for detecting ancient hybridization. Methods Sequencing of a uniparental plastid DNA marker [psbA-trnH(GUG) intergenic spacer] and qualitative and quantitative single nucleotide polymorphism (SNP) genotyping of biparental nuclear ribosomal DNA (nrDNA) markers [LEAFY intron 2 (LFY-i2) and internal transcribed spacer 2 (ITS2)] were used. Key Results Based on the SNP genotyping results, several Platanus accessions show the presence of hybridization/introgression, including some accessions of P. rzedowskii and of P. mexicana var. interior and one of P. mexicana var. mexicana from Oaxaca (= P. oaxacana). Based on haplotype analyses of the psbA-trnH spacer, five haplotypes were detected. The most common of these is present in taxa belonging to P. orientalis, P. racemosa sensu lato, some accessions of P. occidentalis sensu stricto (s.s.) from Texas, P. occidentalis var. palmeri, P. mexicana s.s. and P. rzedowskii. This is highly relevant to genetic relationships with the haplotypes present in P. occidentalis s.s. and P. mexicana var. interior. Conclusions Hybridization and introgression events between lineages ancestral to modern central and eastern North American Platanus species occurred. Plastid haplotypes and qualitative and quantitative SNP genotyping provide information critical for understanding the complex history of Mexican Platanus. Compared with the usual molecular techniques of sub-cloning, sequencing and genotyping, real-time PCR assay is a quick and sensitive technique for analysing complex evolutionary patterns. PMID:23798602

  2. Aurapten, a coumarin with growth inhibition against Leishmania major promastigotes.

    PubMed

    Napolitano, H B; Silva, M; Ellena, J; Rodrigues, B D G; Almeida, A L C; Vieira, P C; Oliva, G; Thiemann, O H

    2004-12-01

    Several natural compounds have been identified for the treatment of leishmaniasis. Among them are some alkaloids, chalcones, lactones, tetralones, and saponins. The new compound reported here, 7-geranyloxycoumarin, called aurapten, belongs to the chemical class of the coumarins and has a molecular weight of 298.37. The compound was extracted from the Rutaceae species Esenbeckia febrifuga and was purified from a hexane extract starting from 407.7 g of dried leaves and followed by four silica gel chromatographic fractionation steps using different solvents as the mobile phase. The resulting compound (47 mg) of shows significant growth inhibition with an LD50 of 30 microM against the tropical parasite Leishmania major, which causes severe clinical manifestations in humans and is endemic in the tropical and subtropical regions. In the present study, we investigated the atomic structure of aurapten in order to determine the existence of common structural motifs that might be related to other coumarins and potentially to other identified inhibitors of Leishmania growth and viability. This compound has a comparable inhibitory activity of other isolated molecules. The aurapten is a planar molecule constituted of an aromatic system with electron delocalization. A hydrophobic side chain consisting of ten carbon atoms with two double bonds and negative density has been identified and may be relevant for further compound synthesis. PMID:15558191

  3. Leishmania spp. Epidemiology of Canine Leishmaniasis in the Yucatan Peninsula

    PubMed Central

    López-Céspedes, A.; Longoni, S. S.; Sauri-Arceo, C. H.; Sánchez-Moreno, M.; Rodríguez-Vivas, R. I.; Escobedo-Ortegón, F. J.; Barrera-Pérez, M. A.; Bolio-González, M. E.; Marín, C.

    2012-01-01

    Canine Leishmaniasis is widespread in various Mexican states, where different species of Leishmania have been isolated from dogs. In the present study, we describe the detection of L. braziliensis, L. infantum, and L. mexicana in serum of dogs from the states of Yucatan and Quintana Roo in the Yucatan Peninsula (Mexico). A total of 412 sera were analyzed by ELISA using the total extract of the parasite and the iron superoxide dismutase excreted by different trypanosomatids as antigens. We found the prevalence of L. braziliensis to be 7.52%, L. infantum to be 6.07%, and L. mexicana to be 20.63%, in the dog population studied. The results obtained with ELISA using iron superoxide dismutase as the antigen were confirmed by western blot analysis with its greater sensitivity, and the agreement between the two techniques was very high. PMID:22927792

  4. A case report and literature review of "Chiclero's ulcer".

    PubMed

    Blaylock, Jason M; Wortmann, Glenn W

    2012-09-01

    An 86-year-old man with history of travel to Guatemala presented with a 4-month history of an enlarging ulcerative lesion on his right ear. After several weeks of empiric treatment for otitis externa, histopathology, culture, and PCR analysis of a biopsy specimen confirmed the diagnosis of localized cutaneous leishmaniasis secondary to Leishmania mexicana. Known as "Chiclero's ulcer" in southeast Mexico and Latin America, this unique presentation of cutaneous leishmaniasis is caused mainly by the L. mexicana complex. Infection results in a single ulcerative lesion, most commonly involving the ear pinna, without a tendency for cutaneous metastasis, lymphatic or mucosal involvement. The majority of cases of "Chiclero's ulcer" spontaneously re-epithelialize without treatment within 3-9 months. This patient's lesion completely resolved without therapy after 11 months. "Chiclero's ulcer" should be considered in the differential diagnosis of a patient presenting with a chronic ulcerative lesion and history of travel to an endemic area. PMID:23146325

  5. Aetiology of visceral leishmaniasis in Mexico.

    PubMed

    Monroy-Ostria, A; Hernandez-Montes, O; Barker, D C

    2000-03-25

    Two children with visceral leishmaniasis (VL), were studied by DNA analysis. DNA from liver biopsy samples from both patients, was amplified by PCR with broad primers specific for the Leishmania subgenus. DNA from the patient from Chiapas was also amplified with primers specific for the Leismania donovani complex and hybridised with a probe specific for L. donovani complex. The second patient, who is the first reported case of visceral leishmaniasis in the Mexican state of Tabasco, where localised cutaneous leishmaniasis and DCL predominate, had a co-infection with Toxoplasma gondii. The DNA from this patient was not amplified with primers specific for the L. donovani complex, did not hybridise with a probe specific for the L. donovani complex, but did hybridise with kDNA from a Mexican Leishmania mexicana strain used as a probe. We therefore, suggest that members of the L. donovani or L. mexicana complexes cause VL in Mexico. PMID:10708655

  6. Male-biased predation of a cave fish by a giant water bug.

    PubMed

    Tobler, Michael; Franssen, Courtney M; Plath, Martin

    2008-08-01

    Male-biased predation has been described from several epigean species, and in many cases, intrinsic differences between the sexes (such as male ornaments) have been suggested as an explanation. Here we report on male-biased predation of a cave fish (Poecilia mexicana) by an aquatic insect (Belostoma sp.) in a Mexican sulfur cave. P. mexicana use aquatic surface respiration (ASR) to survive in their sulfidic, hypoxic habitat. We found that males typically exhibit more ASR activity than females, which leads to increased exposure to the sit-and-wait predator that catches fish near the water surface. Our finding is novel, because male vulnerability to predation is not directly related to male traits involved in courtship, but rather due to other sexual differences in behavior and ultimately, oxygen demands. PMID:18437336

  7. Geographic Distribution of Leishmania Species in Ecuador Based on the Cytochrome B Gene Sequence Analysis.

    PubMed

    Kato, Hirotomo; Gomez, Eduardo A; Martini-Robles, Luiggi; Muzzio, Jenny; Velez, Lenin; Calvopiña, Manuel; Romero-Alvarez, Daniel; Mimori, Tatsuyuki; Uezato, Hiroshi; Hashiguchi, Yoshihisa

    2016-07-01

    A countrywide epidemiological study was performed to elucidate the current geographic distribution of causative species of cutaneous leishmaniasis (CL) in Ecuador by using FTA card-spotted samples and smear slides as DNA sources. Putative Leishmania in 165 samples collected from patients with CL in 16 provinces of Ecuador were examined at the species level based on the cytochrome b gene sequence analysis. Of these, 125 samples were successfully identified as Leishmania (Viannia) guyanensis, L. (V.) braziliensis, L. (V.) naiffi, L. (V.) lainsoni, and L. (Leishmania) mexicana. Two dominant species, L. (V.) guyanensis and L. (V.) braziliensis, were widely distributed in Pacific coast subtropical and Amazonian tropical areas, respectively. Recently reported L. (V.) naiffi and L. (V.) lainsoni were identified in Amazonian areas, and L. (L.) mexicana was identified in an Andean highland area. Importantly, the present study demonstrated that cases of L. (V.) braziliensis infection are increasing in Pacific coast areas. PMID:27410039

  8. Analysis toward innovative herbal antibacterial and antifungal drugs.

    PubMed

    Dhankhar, Sandeep; Dhankhar, Seema; Kumar, Manish; Ruhil, Sonam; Balhara, Meenakshi; Chhillar, Anil K

    2012-12-01

    The antimicrobial activities of four medicinal plants Argemona mexicana, Achyranthes aspera, Catharanthus roseus, and Syzygium cumini were evaluated against Escherichia coli, Vibrio cholerae, Klebsiella pneumoniae, Proteus vulgaris, Bacillus subtilis, Salmonella typhi and three Aspergillus species. Extracts from Achyranthes aspera and Catharanthus roseus showed the highest antimicrobial potential (MIC 0.375-0.750 mg/ml) while extract from Argemona mexicana and Syzygium cumini, showed less activity. In disc diffusion assay, only eight out of twenty extracts showed antimicrobial activity at a concentration of 25.0 μg/ disc. The GCMS investigation reveals the existence of 2-bornanone; 1, 2-benzenedicarboxylic acid, bis (2-methylpropyl) ester; hexadecanoic acid, methyl ester and hexatriacontane in water extract fraction of C. roseus. The present research article provides a review of some medicinal plants incorporating antimicrobial drugs, together with recent advances in emerging therapeutics in clinical development and related patents for exploitation of herbal medicine. PMID:23072646

  9. Geographic Distribution of Leishmania Species in Ecuador Based on the Cytochrome B Gene Sequence Analysis

    PubMed Central

    Kato, Hirotomo; Gomez, Eduardo A.; Martini-Robles, Luiggi; Muzzio, Jenny; Velez, Lenin; Calvopiña, Manuel; Romero-Alvarez, Daniel; Mimori, Tatsuyuki; Uezato, Hiroshi; Hashiguchi, Yoshihisa

    2016-01-01

    A countrywide epidemiological study was performed to elucidate the current geographic distribution of causative species of cutaneous leishmaniasis (CL) in Ecuador by using FTA card-spotted samples and smear slides as DNA sources. Putative Leishmania in 165 samples collected from patients with CL in 16 provinces of Ecuador were examined at the species level based on the cytochrome b gene sequence analysis. Of these, 125 samples were successfully identified as Leishmania (Viannia) guyanensis, L. (V.) braziliensis, L. (V.) naiffi, L. (V.) lainsoni, and L. (Leishmania) mexicana. Two dominant species, L. (V.) guyanensis and L. (V.) braziliensis, were widely distributed in Pacific coast subtropical and Amazonian tropical areas, respectively. Recently reported L. (V.) naiffi and L. (V.) lainsoni were identified in Amazonian areas, and L. (L.) mexicana was identified in an Andean highland area. Importantly, the present study demonstrated that cases of L. (V.) braziliensis infection are increasing in Pacific coast areas. PMID:27410039

  10. Male-biased predation of a cave fish by a giant water bug

    NASA Astrophysics Data System (ADS)

    Tobler, Michael; Franssen, Courtney M.; Plath, Martin

    2008-08-01

    Male-biased predation has been described from several epigean species, and in many cases, intrinsic differences between the sexes (such as male ornaments) have been suggested as an explanation. Here we report on male-biased predation of a cave fish ( Poecilia mexicana) by an aquatic insect ( Belostoma sp.) in a Mexican sulfur cave. P. mexicana use aquatic surface respiration (ASR) to survive in their sulfidic, hypoxic habitat. We found that males typically exhibit more ASR activity than females, which leads to increased exposure to the sit-and-wait predator that catches fish near the water surface. Our finding is novel, because male vulnerability to predation is not directly related to male traits involved in courtship, but rather due to other sexual differences in behavior and ultimately, oxygen demands.

  11. Male mate choice and sperm allocation in a sexual/asexual mating complex of Poecilia (Poeciliidae, Teleostei)

    PubMed Central

    Schlupp, I; Plath, M

    2005-01-01

    Male mate choice is critical for understanding the evolution and maintenance of sexual/asexual mating complexes involving sperm-dependent, gynogenetic species. Amazon mollies (Poecilia formosa) require sperm to trigger embryogenesis, but the males (e.g. Poecilia mexicana) do not contribute genes. Males benefit from mating with Amazon mollies, because such matings make males more attractive to conspecific females, but they might control the cost of such matings by providing less sperm to Amazon mollies. We examined this at the behavioural and sperm levels. P. mexicana males preferred to mate with, and transferred more sperm to conspecific females. However, if males mated with P. formosa, sperm was readily transferred. This underscores the importance of male choice in this system. PMID:17148157

  12. Coastal habitat degradation and green sea turtle diets in Southeastern Brazil.

    USGS Publications Warehouse

    Santos, Robson G.; Martins, Agnaldo Silva; Farias, Julyana da Nobrega; Horta, Antunes Paulo; Pinheiro, Hudson Tercio; Baptistotte, Cecilia; Seminoff, Jeffrey A.; Balazs, George H.; Work, Thierry M.

    2011-01-01

    To show the influence of coastal habitat degradation on the availability of food for green turtles (Chelonia mydas), we assessed the dietary preferences and macroalgae community at a feeding area in a highly urbanized region. The area showed low species richness and was classified as degraded. We examined stomach contents of 15 dead stranded turtles (CCL = 44.0 cm (SD 6.7 cm)). The diet was composed primarily of green algae Ulva spp. (83.6%). In contrast, the macroalgae community was dominated by the green alga Caulerpa mexicana. We found a selection for red algae, seagrass and Ulva spp., and avoidance for C. mexicana and brown alga Dictyopteris delicatula. The low diversity of available food items, possibly a result of environmental degradation, likely contributed to the low dietary diversity. The nutritional implications of this restricted diet are unclear.

  13. New species of Cerambycinae from the Neotropical Region, and nomen novum for Anelaphus maculatus Galileo, Martins, and Santos-Silva, 2014 (Elaphidiini).

    PubMed

    Galileo, Maria Helena M; Martins, Ubirajara R; Santos-Silva, Antonio

    2015-01-01

    Six new species and one new genus are described: Criodion spinosum sp. nov. (Cerambycini), from Bolivia; Eburodacrys wappesi sp. nov. and Eburodacrys skillmani sp. nov. (Eburiini), from Bolivia; Eupempelus rileyorum sp. nov. (Heteropsini) from Panama; Sphalloeme mexicana sp. nov. (Oemini), from Mexico; Wappesoeme camiri sp. nov., new genus (Oemini), from Bolivia. Wappesoeme, Eburodacrys wappesi, E. skillmani, Eupempelus rileyorum, and Criodion spinosum are included in previously published keys. Anelaphus erakyra nomen novum for A. maculatus Galileo et al., 2014 is established. PMID:26250192

  14. Isoenzyme characterization of Leishmania isolated from human cases with localized cutaneous leishmaniasis from the State of Campeche, Yucatan Peninsula, Mexico.

    PubMed

    Canto-Lara, S B; Cardenas-Maruffo, M F; Vargas-Gonzalez, A; Andrade-Narvaez, F

    1998-04-01

    Seventy-five isolates from the State of Campeche, Mexico, an area endemic for localized cutaneous leishmaniasis (LCL), were characterized by isoenzyme markers (glucose phosphate isomerase, mannose phospate isomerase, nucleoside hydrolase, phosphoglucomutase, 6-phosphogluconate dehydrogenase, and glucose-6-phosphate dehydrogenase). Seventy (93.3%) were identified as Leishmania (Leishmania) mexicana and 5 (6.7%) as L. (Viannia) braziliensis. This is the first report of authochthonus human LCL caused by L. (V.) braziliensis in the State of Campeche, Yucatan Peninsula, Mexico. PMID:9574789

  15. Inner Ear Morphology in the Atlantic Molly Poecilia mexicana—First Detailed Microanatomical Study of the Inner Ear of a Cyprinodontiform Species

    PubMed Central

    Schulz-Mirbach, Tanja; Heß, Martin; Plath, Martin

    2011-01-01

    Background Fishes show an amazing diversity in hearing abilities, inner ear structures, and otolith morphology. Inner ear morphology, however, has not yet been investigated in detail in any member of the diverse order Cyprinodontiformes. We, therefore, studied the inner ear of the cyprinodontiform freshwater fish Poecilia mexicana by analyzing the position of otoliths in situ, investigating the 3D structure of sensory epithelia, and examining the orientation patterns of ciliary bundles of the sensory hair cells, while combining μ-CT analyses, scanning electron microscopy, and immunocytochemical methods. P. mexicana occurs in different ecotypes, enabling us to study the intra-specific variability (on a qualitative basis) of fish from regular surface streams, and the Cueva del Azufre, a sulfidic cave in southern Mexico. Results The inner ear of Poecilia mexicana displays a combination of several remarkable features. The utricle is connected rostrally instead of dorso-rostrally to the saccule, and the macula sacculi, therefore, is very close to the utricle. Moreover, the macula sacculi possesses dorsal and ventral bulges. The two studied ecotypes of P. mexicana showed variation mainly in the shape and curvature of the macula lagenae, in the curvature of the macula sacculi, and in the thickness of the otolithic membrane. Conclusions Our study for the first time provides detailed insights into the auditory periphery of a cyprinodontiform inner ear and thus serves a basis—especially with regard to the application of 3D techniques—for further research on structure-function relationships of inner ears within the species-rich order Cyprinodontiformes. We suggest that other poeciliid taxa, or even other non-poeciliid cyprinodontiforms, may display similar inner ear morphologies as described here. PMID:22110746

  16. Mexican species of the genus Stethantyx Townes (Hymenoptera, Ichneumonidae, Tersilochinae)

    PubMed Central

    Khalaim, Andrey I.; Ruíz-Cancino, Enrique

    2013-01-01

    Abstract Six species of the genus Stethantyx Townes are found to occur in Mexico. One species, S. mexicana sp. n., is described as new, and four recently described Neotropical species, S. alajuela Khalaim & Broad, S. heredia Khalaim & Broad, S. osa Khalaim & Broad and S. sanjosea Khalaim & Broad, are new records from Mexico. A key to species of Stethantyx occurring in Mexico is provided. PMID:24363592

  17. Sequence-based molecular phylogenetics and phylogeography of the American box turtles (Terrapene spp.) with support from DNA barcoding.

    PubMed

    Martin, Bradley T; Bernstein, Neil P; Birkhead, Roger D; Koukl, Jim F; Mussmann, Steven M; Placyk, John S

    2013-07-01

    The classification of the American box turtles (Terrapene spp.) has remained enigmatic to systematists. Previous comprehensive phylogenetic studies focused primarily on morphology. The goal of this study was to re-assess the classification of Terrapene spp. by obtaining DNA sequence data from a broad geographic range and from all four recognized species and 11 subspecies within the genus. Tissue samples were obtained for all taxa except for Terrapene nelsoni klauberi. DNA was extracted, and the mitochondrial DNA (mtDNA) cytochrome b (Cytb) and nuclear DNA (nucDNA) glyceraldehyde-3-phosphate-dehydrogenase (GAPD) genes were amplified via polymerase chain reaction and sequenced. In addition, the mtDNA gene commonly used for DNA barcoding (cytochrome oxidase c subunit I; COI) was amplified and sequenced to calculate pairwise percent DNA sequence divergence comparisons for each Terrapene taxon. The sequence data were analyzed using maximum likelihood and Bayesian phylogenetic inference, a molecular clock, AMOVAs, SAMOVAs, haplotype networks, and pairwise percent sequence divergence comparisons. Terrapene carolina mexicana and T. c. yucatana formed a monophyletic clade with T. c. triunguis, and this clade was paraphyletic to the rest of T. carolina. Terrapene ornata ornata and T. o. luteola lacked distinction phylogenetically, and Terrapene nelsoni was confirmed to be the sister taxon of T. ornata. Terrapene c. major, T. c. bauri, and Terrapene coahuila were not well resolved for some of the analyses. The DNA barcoding results indicated that all taxa were different species (>2% sequence divergence) except for T. c. triunguis - T. c. mexicana and T. o. ornata - T. o. luteola. The results suggest that T. c. triunguis should be elevated to species status (Terrapene mexicana), and mexicana and yucatana should be included in this group as subspecies. In addition, T. o. ornata and T. o. luteola should not be considered separate subspecies. The DNA barcoding data support these

  18. Chlamydomonas sajao nov. sp. (Chlorophyta, Volvocales)

    NASA Astrophysics Data System (ADS)

    Lewin, Ralph A.

    1984-06-01

    A new species of Chlamydomonas, namely, C. sajao nov. sp. of the Volvocales, Chlorophyta was isolated from a duckweed growing near a ricefield in the vicinity of Guangzhou, China. This interesting unicellular green alga, similar to C. mexicana from Mexico, secretes quantities of extracellular mucilaginous polysaccharides, and may be employed in improving soil quality. The new species resembles C. waldenburgensis Moewus in most characteristics but differs in three important features.

  19. Behavioral response of two species of stingless bees and the honey bee (Hymenoptera: Apidae) to GF-120.

    PubMed

    Gómez-Escobar, Enoc; Liedo, Pablo; Montoya, Pablo; Vandame, Rémy; Sánchez, Daniel

    2014-08-01

    We present the results of evaluating the response of three species of bees, Trigona fulviventris (Guérin), Scaptotrigona mexicana (Guérin-Meneville), and Apis mellifera (L.), to food sources baited with the toxic bait GF-120 (NF Naturalyte), a spinosad-based bait exclusively used to manage fruit flies. Groups of foragers were trained to collect honey and water from a feeder located 50 m from the colonies. Once a sufficient number of foragers were observed at the experimental location, the training feeder was changed to two or three feeders that offered either honey and water, GF-120, Captor (hydrolyzed protein), GF-120 and honey (4:6), or Captor and honey (1:19). T fulviventris and S. mexicana rarely visited GF-120, Captor, or their mixtures with honey, while approximately 28.5 and 1.5% of A. mellifera foragers visited the GF-120 and honey and Captor and honey mixtures, respectively. Our results show that GF-120 clearly repels T. fulviventris and S. mexicana, whereas for A. mellifera, repellence is not as marked when GF-120 is combined with highly nutritious substances like honey. PMID:25195434

  20. Complementary effect of natural and sexual selection against immigrants maintains differentiation between locally adapted fish

    NASA Astrophysics Data System (ADS)

    Plath, Martin; Riesch, Rüdiger; Oranth, Alexandra; Dzienko, Justina; Karau, Nora; Schießl, Angela; Stadler, Stefan; Wigh, Adriana; Zimmer, Claudia; Arias-Rodriguez, Lenin; Schlupp, Ingo; Tobler, Michael

    2010-08-01

    Adaptation to ecologically heterogeneous environments can drive speciation. But what mechanisms maintain reproductive isolation among locally adapted populations? Using poeciliid fishes in a system with naturally occurring toxic hydrogen sulfide, we show that (a) fish from non-sulfidic sites ( Poecilia mexicana) show high mortality (95 %) after 24 h when exposed to the toxicant, while locally adapted fish from sulfidic sites ( Poecilia sulphuraria) experience low mortality (13 %) when transferred to non-sulfidic water. (b) Mate choice tests revealed that P. mexicana females exhibit a preference for conspecific males in non-sulfidic water, but not in sulfidic water, whereas P. sulphuraria females never showed a preference. Increased costs of mate choice in sulfidic, hypoxic water, and the lack of selection for reinforcement due to the low survival of P. mexicana may explain the absence of a preference in P. sulphuraria females. Taken together, our study may be the first to demonstrate independent—but complementary—effects of natural and sexual selection against immigrants maintaining differentiation between locally adapted fish populations.

  1. Phylogeny of the genus Pistacia as determined from analysis of the chloroplast genome.

    PubMed

    Parfitt, D E; Badenes, M L

    1997-07-22

    Classification within the genus Pistacia has been based on leaf morphology and geographical distribution. Molecular genetic tools (PCR amplification followed by restriction analysis of a 3.2-kb region of variable chloroplast DNA, and restriction fragment length polymorphism analysis of the Pistacia cpDNA with tobacco chloroplast DNA probes) provided a new set of variables to study the phylogenetic relationships of 10 Pistacia species. Both parsimony and cluster analyses were used to divide the genus into two major groups. P. vera was determined to be the least derived species. P. weinmannifolia, an Asian species, is most closely related to P. texana and P. mexicana, New World species. These three species share a common origin, suggesting that a common ancestor of P. texana and P. mexicana originated in Asia. P. integerrima and P. chinensis were shown to be distinct whereas the pairs of species were monophyletic within each of two tertiary groups, P. vera:P. khinjuk and P. mexicana:P. texana. An evolutionary trend from large to small nuts and leaves with few, large leaflets to many, small leaflets was supported. The genus Pistacia was shown to have a low chloroplast DNA mutation rate: 0.05-0.16 times that expected of annual plants. PMID:9223300

  2. At the foot of the shrew: manus morphology distinguishes closely-related Cryptotis goodwini and Cryptotis griseoventris (Mammalia: Soricidae) in Central America

    USGS Publications Warehouse

    Woodman, Neal; Stephens, Ryan B.

    2010-01-01

    Small-eared shrews (Mammalia, Soricidae) of the New World genus Cryptotis are distributed from eastern North America to the northern Andes of South America. One well-defined clade in this genus is the Central American Cryptotis mexicana group, whose members are set off from other species in the genus by their variably broader fore feet and more elongate and broadened fore claws. Two species in the C. mexicana group, Cryptotis goodwini Jackson and Cryptotis griseoventris Jackson, inhabit highlands in Guatemala and southern Mexico and are presumed to be sister species whose primary distinguishing feature is the larger body size of C. goodwini. To better characterize these species and confirm the identification of recently-collected specimens, we obtained digital X-ray images of the manus from large series of dried skins of both species. Measurements of the metacarpals and phalanges successfully separated most specimens of C. goodwini and C. griseoventris. These measurements also show that the fore feet of C. griseoventris from Chiapas, Mexico, are morphologically distinct from those of members of the species inhabiting Guatemala. Univariate, bivariate, and multivariate analyses indicate that fore foot characters are more conservative within species of the C. mexicana group than are cranio-mandibular characters. Patterns of evolution of fore foot characters that superficially appear to be linear gradations are actually more complex, illustrating individual evolutionary trajectories.

  3. Leishmania chitinase facilitates colonization of sand fly vectors and enhances transmission to mice

    PubMed Central

    Rogers, Matthew E; Hajmová, Martina; Joshi, Manju B; Sadlova, Jovana; Dwyer, Dennis M; Volf, Petr; Bates, Paul A

    2008-01-01

    Chitinases of trypanosomatid parasites have been proposed to fulfil various roles in their blood-feeding arthropod vectors but so far none have been directly tested using a molecular approach. We characterized the ability of Leishmania mexicana episomally transfected with LmexCht1 (the L. mexicana chitinase gene) to survive and grow within the permissive sand fly vector, Lutzomyia longipalpis. Compared with control plasmid transfectants, the overexpression of chitinase was found to increase the average number of parasites per sand fly and accelerate the escape of parasites from the peritrophic matrix-enclosed blood meal as revealed by earlier arrival at the stomodeal valve. Such flies also exhibited increased damage to the structure of the stomodeal valve, which may facilitate transmission by regurgitation. When exposed individually to BALB/c mice, those flies with chitinase-overexpressing parasites spent on average 2.4–2.5 times longer in contact with their host during feeding, compared with flies with control infections. Furthermore, the lesions that resulted from these single fly bite infections were both significantly larger and with higher final parasite burdens than controls. These data show that chitinase is a multifunctional virulence factor for L. mexicana which assists its survival in Lu. longipalpis. Specifically, this enzyme enables the parasites to colonize the anterior midgut of the sand fly more quickly, modify the sand fly stomodeal valve and affect its blood feeding, all of which combine to enhance transmission. PMID:18284631

  4. Molecular Phylogeny and Taxonomy of Yamadazyma dushanensis f.a., sp. nov., a Cellobiose-Fermenting Yeast Species from China.

    PubMed

    Wang, Yun; Ren, Yong-Cheng; Li, Ying; Hui, Feng-Li

    2015-08-01

    Three yeast strains of Yamadazyma dushanensis f.a., sp. nov. were isolated from rotten wood samples collected in the Dushan Forest Park, Nanyang, Henan Province, China. Sequence analyses of the D1/D2 domains of the large subunit rRNA gene and the internal transcribed spacer (ITS) regions revealed that this new species is located in the Yamadazyma clade (Debaryomycetaceae and Saccharomycetales), with three closely related species, namely, Yamadazyma terventina, Yamadazyma mexicana and Candida trypodendroni. The novel species differed from these three described species by 5-6 nt substitutions in the D1/D2 sequences. However, the ITS sequences of the new species were quite divergent from those of Y. terventina, Y. mexicana and C. trypodendroni with 12-18 nt substitutions. This new yeast species could assimilate cellobiose and other compounds related to rotting wood. The fermentation of cellobiose in Durham tubes was observed for the strains of this new yeast. The new species could also be distinguished from its closely related species, Y. terventina, Y. mexicana and C. trypodendroni, based on a number of morphological and physiological characteristics. The type strain is Y. dushanensis sp. nov. NYNU 14668 (T) (=CICC 33051(T) = CBS 13914(T)). PMID:26037380

  5. Life on the edge: hydrogen sulfide and the fish communities of a Mexican cave and surrounding waters.

    PubMed

    Tobler, Michael; Schlupp, Ingo; Heubel, Katja U; Riesch, Rüdiger; de León, Francisco J García; Giere, Olav; Plath, Martin

    2006-12-01

    Most eucaryotic organisms classified as living in an extreme habitat are invertebrates. Here we report of a fish living in a Mexican cave (Cueva del Azufre) that is rich in highly toxic H(2)S. We compared the water chemistry and fish communities of the cave and several nearby surface streams. Our study revealed high concentrations of H(2)S in the cave and its outflow (El Azufre). The concentrations of H(2)S reach more than 300 muM inside the cave, which are acutely toxic for most fishes. In both sulfidic habitats, the diversity of fishes was heavily reduced, and Poecilia mexicana was the dominant species indicating that the presence of H(2)S has an all-or-none effect, permitting only few species to survive in sulfidic habitats. Compared to habitats without H(2)S, P. mexicana from the cave and the outflow have a significantly lower body condition. Although there are microhabitats with varying concentrations of H(2)S within the cave, we could not find a higher fish density in areas with lower concentrations of H(2)S. We discuss that P. mexicana is one of the few extremophile vertebrates. Our study supports the idea that extreme habitats lead to an impoverished species diversity. PMID:16788733

  6. In vitro antitrypanosomal activity of ethnopharmacologically selected Beninese plants.

    PubMed

    Hoet, Sara; Opperdoes, Frederik; Brun, Reto; Adjakidjé, Victor; Quetin-Leclercq, Joëlle

    2004-03-01

    The in vitro antitrypanosomal activity of methylene chloride, methanol and aqueous extracts of the leaves and twigs of five plant species traditionally used in Benin for the treatment of sleeping sickness were evaluated on Trypanosoma brucei brucei and their selectivity was analysed on Leishmania mexicana mexicana and J774 macrophage-like murine cells. The results showed that the four most active extracts had MIC values < or =19 microg/ml (Hymenocardia acida twig and leaf, Strychnos spinosa leaf, Trichilia emetica leaf methylene chloride extracts). All these extracts had a lower activity on L. m. mexicana and J774 cells. Determination of the IC50 values of the methylene chloride leaf extracts on two strains of trypanosomes (T. b. brucei and T. b. rhodesiense) and two mammalian cell lines (L6 and J774 cells) showed that all extracts possessed some antitrypanosomal activity with IC50's ranging from 1.5 to 39 microg/ml. All were also toxic to the mammalian cells, but usually with higher IC50's. The only exception was the S. spinosa methylene chloride leaf extract which had no toxicity on J774 cells. Although tannins have been identified in most of the species studied, they could not be detected in the most active extracts, just as alkaloids. The presence of flavonoids and quinones may at least in part explain the observed activities of some of the active extracts. PMID:15036465

  7. Phylogeny of the genus Pistacia as determined from analysis of the chloroplast genome

    PubMed Central

    Parfitt, Dan E.; Badenes, Maria L.

    1997-01-01

    Classification within the genus Pistacia has been based on leaf morphology and geographical distribution. Molecular genetic tools (PCR amplification followed by restriction analysis of a 3.2-kb region of variable chloroplast DNA, and restriction fragment length polymorphism analysis of the Pistacia cpDNA with tobacco chloroplast DNA probes) provided a new set of variables to study the phylogenetic relationships of 10 Pistacia species. Both parsimony and cluster analyses were used to divide the genus into two major groups. P. vera was determined to be the least derived species. P. weinmannifolia, an Asian species, is most closely related to P. texana and P. mexicana, New World species. These three species share a common origin, suggesting that a common ancestor of P. texana and P. mexicana originated in Asia. P. integerrima and P. chinensis were shown to be distinct whereas the pairs of species were monophyletic within each of two tertiary groups, P. vera:P. khinjuk and P. mexicana:P. texana. An evolutionary trend from large to small nuts and leaves with few, large leaflets to many, small leaflets was supported. The genus Pistacia was shown to have a low chloroplast DNA mutation rate: 0.05–0.16 times that expected of annual plants. PMID:9223300

  8. Critical role for phosphoinositide 3-kinase gamma in parasite invasion and disease progression of cutaneous leishmaniasis

    PubMed Central

    Cummings, Hannah E.; Barbi, Joseph; Reville, Patrick; Oghumu, Steve; Zorko, Nicholas; Sarkar, Anasuya; Keiser, Tracy L.; Lu, Bao; Rückle, Thomas; Varikuti, Sanjay; Lezama-Davila, Claudio; Wewers, Mark D.; Whitacre, Caroline; Radzioch, Danuta; Rommel, Christian; Seveau, Stéphanie; Satoskar, Abhay R.

    2012-01-01

    Obligate intracellular pathogens such as Leishmania specifically target host phagocytes for survival and replication. Phosphoinositide 3-kinase γ (PI3Kγ), a member of the class I PI3Ks that is highly expressed by leukocytes, controls cell migration by initiating actin polymerization and cytoskeletal reorganization, which are processes also critical for phagocytosis. In this study, we demonstrate that class IB PI3K, PI3Kγ, plays a critical role in pathogenesis of chronic cutaneous leishmaniasis caused by L. mexicana. Using the isoform-selective PI3Kγ inhibitor, AS-605240 and PI3Kγ gene-deficient mice, we show that selective blockade or deficiency of PI3Kγ significantly enhances resistance against L. mexicana that is associated with a significant suppression of parasite entry into phagocytes and reduction in recruitment of host phagocytes as well as regulatory T cells to the site of infection. Furthermore, we demonstrate that AS-605240 is as effective as the standard antileishmanial drug sodium stibogluconate in treatment of cutaneous leishmaniasis caused by L. mexicana. These findings reveal a unique role for PI3Kγ in Leishmania invasion and establishment of chronic infection, and demonstrate that therapeutic targeting of host pathways involved in establishment of infection may be a viable strategy for treating infections caused by obligate intracellular pathogens such as Leishmania. PMID:22232690

  9. Pi inhibits intracellular accumulation of methylglyoxal in promastigote form of L. donovani.

    PubMed

    Tiwari, Praachi; Verma, Pankaj; Kureel, Amit Kumar; Saini, Sheetal; Rai, Ambak Kumar

    2016-06-01

    Similar to their mammalian counterpart, protozoan parasites including Leishmania donovani detoxify methylglyoxal (MG),(1) a toxic ubiquitous product generated in glycolysis pathway. However, it differs in one or more way(s) from the humans. It is known that MG is eliminated either through glyoxalase (GLO)(2) pathway and/or excreted across the cell membrane. This toxic metabolic by-product is known to be detoxified predominantly by the GLO pathway and excretion across the cell membrane is never considered to be a significant pathway for its detoxification. We have tried to modulate these pathways under various physiological conditions to find ways that may lead to accumulation of MG in L. donovani. Besides targeting the GLO pathway, we intend to understand the mechanism of MG release across the cell membrane and possible ways to inhibit its exclusion from parasites. In our study, it was found that inorganic phosphate (Pi)(3) in the presence of glucose inhibits the production of MG as well as promotes the expulsion of MG from the cell. Moreover, the trivalent form of antimony (Sb(III)) inhibits GLO pathway and thus detoxification of MG. Inhibition of Pi transport, which is a Na(+)/H(+) dependent process, restores the Pi mediated abrogation of MG production. Thus, Sb(III) along with inhibitors of Pi transporter may be a therapeutic advancement for treatment of antimony resistant cases of human visceral leishmaniasis. However, it requires further validation using specific inhibitor(s) of Pi transporter. PMID:27297182

  10. Antiproteolytic and leishmanicidal activity of Coccinia grandis (L.) Voigt leaf extract against Leishmania donovani promastigotes.

    PubMed

    Das, Partha; Paik, Dibyendu; Pramanik, Asmita; De, Tripti; Chakraborti, Tapati

    2015-11-01

    In visceral leishmaniasis (VL), development of alternative safe therapeutic strategy is gaining paramount wherein natural components of plant origin have prominence. We explored Coccinia grandis (L.) Voigt, a medicinal plant known in traditional folk medicine, for its antileishmanial efficacy. SDS-PAGE analysis of the C. grandis leaf extract (Cg-Ex) showed few protein bands about 14-66 kDa among which three (64.8, 55.8 and 15.3 kDa) were identified as serine protease inhibitors by reverse zymography. Since the virulence of Leishmania is also attributed by serine proteases, objective of the present study was to evaluate in vitro antileishmanial activity of Cg-Ex, targeting Leishmania donovani serine protease(s). Inhibition study of Cg-Ex in gelatin-zymogram and spectrophotometric assay revealed its strong inhibitory activity against bovine trypsin rather than chymotrypsin, and also showed significant inhibition of L. donovani serine protease(s). Further, studies with Cg-Ex were extended to estimate its antileishmanial efficacy with half maximal inhibitory concentration (IC50) at 308.0 ± 2.42 μg/ml along with significant morphological alterations. The results have demonstrated the potential of the serine protease inhibitor rich fraction of the C. grandis leaf extract against visceral leishmaniasis. PMID:26669017

  11. Antioxidant and free radical scavenging activities of some fruits.

    PubMed

    Prakash, Dhan; Upadhyay, Garima; Pushpangadan, P; Gupta, Charu

    2011-01-01

    Phenols, a major group of antioxidant phytochemicals, have profound importance due to their biological and free radical scavenging activities. To identify their potential sources extracts of some fruits and their different parts were studied for total phenolic contents (TPC), antioxidant (AOA) and free radical scavenging activities (FRSA). The amount of TPC varied from 10.5 (Carissa carandus, fruit peel) to 343.2 mg/g (Caesalpinia Mexicana, fruits) and AOA from 20.3% (Musa paradisiacal, fruits) to 96.7% (Caesalpinia Mexicana, fruits). Fruits of Caesalpinia Mexicana, Acacia auriculiformis, fruit pericarp green fibres of Cocus nucifera, and fruits of Emblica officinalis were found to have high TPC (73.1-343.2 mg/g) and high AOA (68.5-96.7%). Promising fruits were studied for their FRSA and reducing power (RP) measured by DPPH assay where the fruits of Caesalpinia mexicana, fruit pericarp fibres of Cocus nucifera, fruits of Emblica officinalis showed very low IC50 ranging from 0.009 to 0.016 mg/ml, EC50 from 0.39 to 0.70 mg/mg DPPH and reasonably high values (142.1-256.3) of anti radical power (ARP), indicating their strong FRSA and reducing power (RP) as evident by their low ASE/ml values (0.42-1.08). They also showed better inhibition of lipid peroxidation measured by using ferric thiocyanate assay and by using egg yolk compared to the reference standard quercetin. The ferrous and ferric ion chelating capacity of the promising fruits and their underutilized parts in terms of IC50 varied from 0.12 (Emblica officinalis, fruits) to 2.44 mg/ml (Mangifera indica, Seed kernel) and 0.22 (Caesalpinia Mexicana, fruits) to 2.59 mg/ml (Litchi chinensis, fruit peel) respectively. Fruit pulp, peel and seeds of Litchi chinensis with reasonable amount of phenols (48.3, 43.9, 50.1 mg/ml) showed low ARP (23.5, 38.3, 33.8) and ASE/ml (3.13, 2.18, 2.62) respectively in contrast to Aegle marmelos with comparatively lower phenols (35.1 mg/g) exhibited good ARP (57.4) and RP (1.67 ASE

  12. Effects of harvest on the sustainability and leaf productivity of populations of two palm species in Maya homegardens.

    PubMed

    Martínez-Ballesté, Andrea; Martorell, Carlos

    2015-01-01

    Traditional management practices are usually thought to be sustainable. The Maya manage Sabal (Arecaceae) palms in homegardens, using their leaves for thatching. The sustainability of such production systems depends on the long-term persistence of palm populations, whereas resource availability also depends on the number of leaves on individual palms. We examined how leaf harvest affects Sabal yapa and S. mexicana population growth rates (λ) and leaf production, comparing traditional and alternative harvest regimes in terms of sustainability and productivity. Demographic, harvest and leaf production data were recorded for three years in two homegardens. We used general integral projection models linked to leaf-production models to describe population dynamics and productivity. Harvest had no effect on S. yapa's vital rates or on λ, but it changed the growth rate of individuals of S. mexicana, with a negligible impact on λ. Homegardens affected λ values, reflecting the species' ecological affinities. S. mexicana, introduced from mesic forests, required watering and shade; therefore, its population declined rapidly in the homegarden that lacked both water and shade. The λ of the xerophilic S. yapa was slightly larger without watering than with watering. Palms usually compensated for leaf extraction, causing the number of leaves harvested per individual to increase with harvest intensity. Nevertheless, traditional management is relatively mild, allowing standing leaves to accumulate but reducing the homegarden's yield. Apparently, the Maya do not seek to maximize annual production but to ensure the availability of large numbers of leaves in homegardens. These leaves may then be used when the entire roof of a hut needs to be replaced every few years. PMID:25803029

  13. Effects of Harvest on the Sustainability and Leaf Productivity of Populations of Two Palm Species in Maya Homegardens

    PubMed Central

    Martínez-Ballesté, Andrea; Martorell, Carlos

    2015-01-01

    Traditional management practices are usually thought to be sustainable. The Maya manage Sabal (Arecaceae) palms in homegardens, using their leaves for thatching. The sustainability of such production systems depends on the long-term persistence of palm populations, whereas resource availability also depends on the number of leaves on individual palms. We examined how leaf harvest affects Sabal yapa and S. mexicana population growth rates (λ) and leaf production, comparing traditional and alternative harvest regimes in terms of sustainability and productivity. Demographic, harvest and leaf production data were recorded for three years in two homegardens. We used general integral projection models linked to leaf-production models to describe population dynamics and productivity. Harvest had no effect on S. yapa’s vital rates or on λ, but it changed the growth rate of individuals of S. mexicana, with a negligible impact on λ. Homegardens affected λ values, reflecting the species’ ecological affinities. S. mexicana, introduced from mesic forests, required watering and shade; therefore, its population declined rapidly in the homegarden that lacked both water and shade. The λ of the xerophilic S. yapa was slightly larger without watering than with watering. Palms usually compensated for leaf extraction, causing the number of leaves harvested per individual to increase with harvest intensity. Nevertheless, traditional management is relatively mild, allowing standing leaves to accumulate but reducing the homegarden’s yield. Apparently, the Maya do not seek to maximize annual production but to ensure the availability of large numbers of leaves in homegardens. These leaves may then be used when the entire roof of a hut needs to be replaced every few years. PMID:25803029

  14. Antinociceptive and Anti-Inflammatory Activity from Algae of the Genus Caulerpa

    PubMed Central

    da Matta, Carolina Babosa Brito; de Souza, Éverton Tenório; de Queiroz, Aline Cavalcanti; de Lira, Daysianne Pereira; de Araújo, Morgana Vital; Cavalcante-Silva, Luiz Henrique Agra; de Miranda, George Emmanuel C.; de Araújo-Júnior, João Xavier; Barbosa-Filho, José Maria; de Oliveira Santos, Bárbara Viviana; Alexandre-Moreira, Magna Suzana

    2011-01-01

    Marine natural products have been the focus of discovery for new products of chemical and pharmacological interest. The aim of this study was to evaluate the antinociceptive activity of the methanolic (ME), acetate (AE), hexanic (HE) and chloroform (CE) extracts obtained from Caulerpa mexicana, and ME, CE and HE obtained from Caulerpa sertularioides. These marine algae are found all over the world, mainly in tropical regions. Models such as the writhing test, the hot plate test and formalin-induced nociception test were used to evaluate antinociceptive activity in laboratory mice. In the writhing test, all the extracts were administered orally at a concentration of 100 mg/kg, and induced high peripheral antinociceptive activity, with a reduction in the nociception induced by acetic acid above 65%. In the hot plate test, treatment with extracts from C. sertularioides (100 mg/kg, p.o.) did not significantly increase the latency of response, although the ME, AE and HE from C. mexicana showed activity in this model. This result suggests that these extracts exhibit antinociceptive activity. In the formalin test, it was observed that ME, AE and HE obtained from C. mexicana reduced the effects of formalin in both phases. On the other hand only CE from C. sertularioides induced significant inhibition of the nociceptive response in the first phase. To better assess the potential anti-inflammatory activity of the extracts, the carrageenan-induced peritonitis test was used to test Caulerpa spp. extracts on cell migration into the peritoneal cavity. In this assay, all extracts evaluated were able to significantly inhibit leukocyte migration into the peritoneal cavity in comparison with carrageenan. These data demonstrate that extracts from Caulerpa species elicit pronounced antinociceptive and anti-inflamatory activity against several nociception models. However, pharmacological and chemical studies are continuing in order to characterize the mechanism(s) responsible for the

  15. Morphological and molecular description of eight new species of Gyrodactylus von Nordmann, 1832 (Platyhelminthes: Monogenea) from poeciliid fishes, collected in their natural distribution range in the Gulf of Mexico slope, Mexico.

    PubMed

    García-Vásquez, Adriana; Razo-Mendivil, Ulises; Rubio-Godoy, Miguel

    2015-09-01

    Eight new species of Gyrodactylus are described from Poecilia mexicana, Poeciliopsis gracilis, Pseudoxiphophorus bimaculatus [syn. = Heterandria bimaculata], and Xiphophorus hellerii collected in the Nautla and La Antigua River Basins in Veracruz, and in the Tecolutla River Basin in Puebla, Mexico. Analyzing the morphology of the marginal hooks, Gyrodactylus pseudobullatarudis n. sp. and Gyrodactylus xtachuna n. sp. are both very similar to Gyrodactylus bullatarudis; Gyrodactylus takoke n. sp. resembles Gyrodactylus xalapensis; Gyrodactylus lhkahuili n. sp. is similar to Gyrodactylus jarocho; and both Gyrodactylus microdactylus n. sp. and Gyrodactylus actzu n. sp. are similar to Gyrodactylus poeciliae in that all three species possess extremely short shaft points. A hypothesis of the systematic relationships of the eight new Gyrodactylus species and some of the known gyrodactylids infecting poeciliids was constructed with sequences of the Internal Transcribed Spacers (ITS1 and ITS2) and the 5.8S ribosomal gene of the rRNA. Phylogenetic trees showed that the new and previously described species of Gyrodactylus infecting poeciliid fishes do not form a monophyletic assemblage. Trees also showed that the eight new species described morphologically correspond to well-supported monophyletic groups; and that morphologically similar species are also phylogenetically close. Additionally, we correct previous erroneous records of the presence of Gyrodactylus bullatarudis on wild Poecilia mexicana and Xiphophorus hellerii collected in Mexico, as re-examination of the original specimens indicated that these corresponded to Gyrodactylus pseudobullatarudis n. sp. (infecting Poecilia mexicana and Xiphophorus hellerii) and to Gyrodactylus xtachuna n. sp. (on Xiphophorus hellerii). Finally, given the widespread anthropogenic translocation of poeciliid fishes for the aquarium trade and mosquito control programs, as well as the existence of invasive, feral poeciliid populations

  16. Antimicrobial evaluation of some plants used in Mexican traditional medicine for the treatment of infectious diseases.

    PubMed

    Navarro, V; Villarreal, M L; Rojas, G; Lozoya, X

    1996-09-01

    Twelve methanolic plant extracts from botanical species used in traditional medicine in Morelos, México to cure infectious diseases have been subjected to a screening study to detect potential antimicrobial activity against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans. The antimicrobial activity of the products was evaluated using colonies growing in solid medium, establishing the minimal concentration required to inhibit their in vitro growth (MIC). The results showed that extracts from Eucalyptus globolus Labill, Punica granatum L., Artemisia mexicana Wild., and Bocconia arborea Watt. possess strong in vitro antimicrobial activity against the tested microorganisms. PMID:8887021

  17. The genus Triozocera Pierce, 1909 (Insecta: Strepsiptera: Corioxenidae) in South America.

    PubMed

    Kogan, Marcos; Cook, Jerry L

    2014-01-01

    A new species of Triozocera from the Brazilian Amazon basin was found in a sample of male Strepsiptera from the collection of the Instituto Nacional de Pesquisas da Amazonia (INPA-Manaus, Amazonas, Brazil). Triozocera buehrheimi sp.n. is described and the status of T. paulistana Kogan, 1958, the first strepsipteran described from Brazil, is reviewed, with additional diagnostic characters used to reinstate the species based on comparative analyses to the other three species occurring in southern US, Mexico, and Central America: T. mexicana Pierce, 1909, T. tecpanensis Brailowsky and Márquez, 1974, and T. vernalis Kifune and Brailowsky, 1987. A key to those species is included. PMID:24871713

  18. A revision of the genus Protorthodes McDunnough with descriptions of a new genus and four new species (Lepidoptera, Noctuidae, Noctuinae, Eriopygini)

    PubMed Central

    Lafontaine, J. Donald; Walsh, J. Bruce; Ferris, Clifford D.

    2014-01-01

    Abstract The genus Protorthodes McDunnough is revised to include 15 species including P. ustulata Lafontaine, Walsh & Ferris, sp. n., from southwestern United States, P. texicana Lafontaine, sp. n., from Texas and Mexico, and P. mexicana Lafontaine, sp. n., from Mexico. A new genus, Nudorthodes Lafontaine, Walsh & Ferris, is proposed for two species formerly included in Protorthodes, P. texana (Smith, 1900), comb. n., and P. variabilis (Barnes & McDunnough, 1912), comb. n., and N. molino Lafontaine, Walsh & Ferris, sp. n., described from southern Arizona. A key to species, descriptions, illustrations of adults and genitalia, and distribution maps are included. PMID:25061383

  19. The ALICE Pixel Detector

    NASA Astrophysics Data System (ADS)

    Mercado-Perez, Jorge

    2002-07-01

    The present document is a brief summary of the performed activities during the 2001 Summer Student Programme at CERN under the Scientific Summer at Foreign Laboratories Program organized by the Particles and Fields Division of the Mexican Physical Society (Sociedad Mexicana de Fisica). In this case, the activities were related with the ALICE Pixel Group of the EP-AIT Division, under the supervision of Jeroen van Hunen, research fellow in this group. First, I give an introduction and overview to the ALICE experiment; followed by a description of wafer probing. A brief summary of the test beam that we had from July 13th to July 25th is given as well.

  20. [A critical view about public health in Mexico].

    PubMed

    Tapia-Conyer, Roberto

    2016-01-01

    La salud pública se encuentra en un momento crucial, el cual tenemos que aprovechar para devolver a México el liderazgo que lo distinguió a nivel regional y mundial. Para ello, quienes hemos dedicado esfuerzos profesionales al fortalecimiento de la salud pública mexicana debemos entender el estado que guardan la ciencia, las nuevas tecnologías y los nuevos desarrollos para no solamente evitar el rezago, sino situarnos en la vanguardia, como corresponde a la acreditada tradición médica de nuestro país. PMID:27160628

  1. Receptor-Mediated Drug Delivery to Macrophages in Chemotherapy of Leishmaniasis

    NASA Astrophysics Data System (ADS)

    Mukhopadhyay, Amitabha; Chaudhuri, Gautam; Arora, Sunil K.; Sehgal, Shobha; Basu, Sandip K.

    1989-05-01

    Methotrexate coupled to maleylated bovine serum albumin was taken up efficiently through the ``scavenger'' receptors present on macrophages and led to selective killing of intracellular Leishmania mexicana amazonensis amastigotes in cultured hamster peritoneal macrophages. The drug conjugate was nearly 100 times as effective as free methotrexate in eliminating the intracellular parasites. Furthermore, in a model of experimental cutaneous leishmaniasis in hamsters, the drug conjugate brought about more than 90% reduction in the size of footpad lesions within 11 days. In contrast, the free drug at a similar concentration did not significantly affect lesion size. These studies demonstrate the potential of receptor-mediated drug delivery in the therapy of macrophage-associated diseases.

  2. Is lipophosphoglycan a virulence factor? A surprising diversity between Leishmania species.

    PubMed

    Turco, S J; Späth, G F; Beverley, S M

    2001-05-01

    Lipophosphoglycan is a prominent member of the phosphoglycan-containing surface glycoconjugates of Leishmania. Genetic tests enable confirmation of its role in parasite virulence and permit discrimination between the roles of lipophosphoglycan and related glycoconjugates. When two different lipophosphoglycan biosynthetic genes from Leishmania major were knocked out, there was a clear loss of virulence in several steps of the infectious cycle but, with Leishmania mexicana, no effect on virulence was found. This points to an unexpected diversity in the reliance of Leishmania species on virulence factors, a finding underscored by recent studies showing great diversity in the host response to Leishmania species. PMID:11323305

  3. New World species of Asymphyloptera Collin (Diptera: Empididae: Clinocerinae).

    PubMed

    Sinclair, Bradley J

    2015-01-01

    Eight species are recognized among New World species of Asymphyloptera Collin, including seven new species (A. cajanuma sp. nov. (Ecuador), A. chilensis sp. nov. (Chile), A. chiricahua sp. nov. (USA: Arizona), A. dominica sp. nov. (Dominica), A. havasu sp. nov. (USA: Arizona), A. lutea sp. nov. (Costa Rica) and A. mexicana sp. nov. (Mexico)). The new species are described, male terminalia illustrated, distributions mapped and a key to species is presented. Two additional undescribed species based on single females, are known from Ecuador and Venezuela. PMID:26624767

  4. Molecular Phylogeny Supports Repeated Adaptation to Burrowing within Small-Eared Shrews Genus of Cryptotis (Eulipotyphla, Soricidae)

    PubMed Central

    He, Kai; Woodman, Neal; Boaglio, Sean; Roberts, Mariel; Supekar, Sunjana; Maldonado, Jesús E.

    2015-01-01

    Small-eared shrews of the New World genus Cryptotis (Eulipotyphla, Soricidae) comprise at least 42 species that traditionally have been partitioned among four or more species groups based on morphological characters. The Cryptotis mexicana species group is of particular interest, because its member species inhibit a subtly graded series of forelimb adaptations that appear to correspond to locomotory behaviors that range from more ambulatory to more fossorial. Unfortunately, the evolutionary relationships both among species in the C. mexicana group and among the species groups remain unclear. To better understand the phylogeny of this group of shrews, we sequenced two mitochondrial and two nuclear genes. To help interpret the pattern and direction of morphological changes, we also generated a matrix of morphological characters focused on the evolutionarily plastic humerus. We found significant discordant between the resulting molecular and morphological trees, suggesting considerable convergence in the evolution of the humerus. Our results indicate that adaptations for increased burrowing ability evolved repeatedly within the genus Cryptotis. PMID:26489020

  5. Genomic variation in recently collected maize landraces from Mexico.

    PubMed

    Arteaga, María Clara; Moreno-Letelier, Alejandra; Mastretta-Yanes, Alicia; Vázquez-Lobo, Alejandra; Breña-Ochoa, Alejandra; Moreno-Estrada, Andrés; Eguiarte, Luis E; Piñero, Daniel

    2016-03-01

    The present dataset comprises 36,931 SNPs genotyped in 46 maize landraces native to Mexico as well as the teosinte subspecies Zea maiz ssp. parviglumis and ssp. mexicana. These landraces were collected directly from farmers mostly between 2006 and 2010. We accompany these data with a short description of the variation within each landrace, as well as maps, principal component analyses and neighbor joining trees showing the distribution of the genetic diversity relative to landrace, geographical features and maize biogeography. High levels of genetic variation were detected for the maize landraces (H E = 0.234 to 0.318 (mean 0.311), while slightly lower levels were detected in Zea m. mexicana and Zea m. parviglumis (H E = 0.262 and 0.234, respectively). The distribution of genetic variation was better explained by environmental variables given by the interaction of altitude and latitude than by landrace identity. This dataset is a follow up product of the Global Native Maize Project, an initiative to update the data on Mexican maize landraces and their wild relatives, and to generate information that is necessary for implementing the Mexican Biosafety Law. PMID:26981357

  6. Characterization of Leishmania (Leishmania) waltoni n.sp. (Kinetoplastida: Trypanosomatidae), the Parasite Responsible for Diffuse Cutaneous Leishmaniasis in the Dominican Republic.

    PubMed

    Shaw, Jeffrey; Pratlong, Francine; Floeter-Winter, Lucile; Ishikawa, Edna; El Baidouri, Fouad; Ravel, Christophe; Dedet, Jean-Pierre

    2015-09-01

    Leishmania parasites isolated, between 1979 and 1988 by the late Bryce Walton, from Dominican Republic (DR) patients with diffuse cutaneous leishmaniasis, were characterized using a panel of 12 isoenzymes, 23 monoclonal antibodies, small subunit ribosomal DNA (SSu rDNA), and multilocus sequence analysis (MLSA). The isoenzyme and monoclonal antibody profiles and the MLSA results showed that the Dominican Republic parasites were distinct from other described Leishmania species. This new species belongs to the mexicana complex, which is distributed in central and parts of northern South America. It is suggested that the parasites uniqueness from other members of the mexicana complex is related to it being isolated on an island for millions of years. If Leishmania (Leishmania) waltoni fails to adapt to some imported mammal, such as the house rat, it will be the only Leishmania to be classified as an endangered species. The excessive destruction of habitats on Hispaniola threatens the survival of its vectors and presumed natural reservoirs, such as the rodent hutias and the small insectivorous mammal solenodon. The concept of Leishmania species is discussed in the light of recent evaluations on criteria for defining bacterial species. PMID:26149864

  7. Molecular Phylogeny Supports Repeated Adaptation to Burrowing within Small-Eared Shrews Genus of Cryptotis (Eulipotyphla, Soricidae).

    PubMed

    He, Kai; Woodman, Neal; Boaglio, Sean; Roberts, Mariel; Supekar, Sunjana; Maldonado, Jesús E

    2015-01-01

    Small-eared shrews of the New World genus Cryptotis (Eulipotyphla, Soricidae) comprise at least 42 species that traditionally have been partitioned among four or more species groups based on morphological characters. The Cryptotis mexicana species group is of particular interest, because its member species inhibit a subtly graded series of forelimb adaptations that appear to correspond to locomotory behaviors that range from more ambulatory to more fossorial. Unfortunately, the evolutionary relationships both among species in the C. mexicana group and among the species groups remain unclear. To better understand the phylogeny of this group of shrews, we sequenced two mitochondrial and two nuclear genes. To help interpret the pattern and direction of morphological changes, we also generated a matrix of morphological characters focused on the evolutionarily plastic humerus. We found significant discordant between the resulting molecular and morphological trees, suggesting considerable convergence in the evolution of the humerus. Our results indicate that adaptations for increased burrowing ability evolved repeatedly within the genus Cryptotis. PMID:26489020

  8. Characterization of Leishmania (Leishmania) waltoni n.sp. (Kinetoplastida: Trypanosomatidae), the Parasite Responsible for Diffuse Cutaneous Leishmaniasis in the Dominican Republic

    PubMed Central

    Shaw, Jeffrey; Pratlong, Francine; Floeter-Winter, Lucile; Ishikawa, Edna; El Baidouri, Fouad; Ravel, Christophe; Dedet, Jean-Pierre

    2015-01-01

    Leishmania parasites isolated, between 1979 and 1988 by the late Bryce Walton, from Dominican Republic (DR) patients with diffuse cutaneous leishmaniasis, were characterized using a panel of 12 isoenzymes, 23 monoclonal antibodies, small subunit ribosomal DNA (SSu rDNA), and multilocus sequence analysis (MLSA). The isoenzyme and monoclonal antibody profiles and the MLSA results showed that the Dominican Republic parasites were distinct from other described Leishmania species. This new species belongs to the mexicana complex, which is distributed in central and parts of northern South America. It is suggested that the parasites uniqueness from other members of the mexicana complex is related to it being isolated on an island for millions of years. If Leishmania (Leishmania) waltoni fails to adapt to some imported mammal, such as the house rat, it will be the only Leishmania to be classified as an endangered species. The excessive destruction of habitats on Hispaniola threatens the survival of its vectors and presumed natural reservoirs, such as the rodent hutias and the small insectivorous mammal solenodon. The concept of Leishmania species is discussed in the light of recent evaluations on criteria for defining bacterial species. PMID:26149864

  9. Egg Load Decreases Mobility and Increases Predation Risk in Female Black-Horned Tree Crickets (Oecanthus nigricornis)

    PubMed Central

    Ercit, Kyla; Martinez-Novoa, Andrew; Gwynne, Darryl T.

    2014-01-01

    Female-biased predation is an uncommon phenomenon in nature since males of many species take on riskier behaviours to gain more mates. Several species of sphecid wasps have been observed taking more female than male prey, and it is not fully understood why. The solitary sphecid Isodontia mexicana catches more adult female tree cricket (Oecanthus nigricornis) prey. Previous work has shown that, although female tree crickets are larger and thus likely to be more valuable as prey than males, body size alone cannot fully explain why wasps take more females. We tested the hypothesis that wasps catch adult female tree crickets more often because bearing eggs impedes a female’s ability to escape predation. We compared female survivors to prey of I. mexicana, and found that females carrying more eggs were significantly more likely to be caught by wasps, regardless of their body size and jumping leg mass. We also conducted laboratory experiments where females’ jumping responses to a simulated attack were measured and compared to her egg load and morphology. We found a significant negative relationship between egg load and jumping ability, and a positive relationship between body size and jumping ability. These findings support the hypothesis that ovarian eggs are a physical handicap that contributes to female-biased predation in this system. Predation on the most fecund females may have ecological-evolutionary consequences such as collapse of prey populations or selection for alternate life history strategies and behaviours. PMID:25330090

  10. Isolation and Characterization of Some Phytochemicals from Indian Traditional Plants

    PubMed Central

    Srivastava, Neeharika; Chauhan, Aishwarya Singh; Sharma, Bechan

    2012-01-01

    The present study was designed to evaluate relative contribution of different polyphenols (total phenolics, flavonoids, flavonols) and their antioxidants activities in aqueous extracts of different parts of some plants; Argemone mexicana, Datura metel, Calotropis procera, Thevetia peruviana, and Cannabis sativa. The antioxidants (total phenolics, flavonoids, flavones) were determined by chemical methods. The antioxidant capacities of these extracts were evaluated by FRAP assay. The results demonstrated that phenolic content was maximally present in leaves of T. peruviana. This plant exhibited minimum phenolic content in its flower as compared to other plants. The flower of D. metel contained maximum phenolic content. The flavonoids were present in highest quantity in leaves of C. procera while T. peruviana flowers showed maximum flavonoid content. The fruits of C. sativa contained maximum quantity of flavonoid as compared to other plants tested. The flower extract of C. sativa possessed highest FRAP value followed by A. mexicana and fruit of C. procera. The values of ratios of different polyphenolic compounds present in plant extracts indicated that flower of D. metel contained maximum total flavonoids and minimum phenolics. These results suggested that levels of total phenolics, flavonoids and their FRAP indices exhibited specificity to different plants and their parts. PMID:23304524

  11. Antibody response against a Leishmania donovani amastigote-stage-specific protein in patients with visceral leishmaniasis.

    PubMed Central

    Ghedin, E; Zhang, W W; Charest, H; Sundar, S; Kenney, R T; Matlashewski, G

    1997-01-01

    The antibody response against an amastigote-specific protein (A2) from Leishmania donovani was investigated. Sera from patients with trypanosomiasis and various forms of leishmaniasis were screened for anti-A2 antibodies. Sera from patients infected only with L. donovani or Leishmania mexicana specifically recognized the A2 recombinant protein. These results were consistent with karyotype analyses which revealed that the A2 gene is conserved in L. donovani and L. mexicana strains. The potential of this antigen in diagnosis was further explored by screening a series of sera obtained from patients in regions of the Sudan and India where L. donovani is endemic. The prevalence of anti-A2 antibodies was determined by Western blotting for all samples. Enzyme-linked immunosorbent assay (ELISA) and an immunoprecipitation assay were also performed on some of the samples. Anti-A2 antibodies were detected by ELISA in 82 and 60% of the samples from individuals with active visceral leishmaniasis (kala-azar) from the Sudan and India, respectively, while the immunoprecipitation assay detected the antibodies in 92% of the samples from India. These data suggest that the A2 protein may be a useful diagnostic antigen for visceral leishmaniasis. PMID:9302200

  12. Reproductive characteristics of a captive colony of big-eared climbing rats (Ototylomys phyllotis).

    PubMed

    Itzá-Ortiz, Mateo F; Van Wynsberghe, Nicole R; Sosa-Bibiano, Erika I; Andrade-Narváez, Fernando J

    2011-08-01

    The authors analyzed the breeding characteristics of a colony of Ototylomys phyllotis (big-eared climbing rat) from Campeche, México, that was bred in captivity for 6 y. The big-eared climbing rat is a reservoir of Leishmania (Leishmania) mexicana, a causal agent of localized cutaneous leishmaniasis on the Yucatán Peninsula. The colony had been established to facilitate studies analyzing the effectiveness of O. phyllotis as an experimental model for L. (L.) mexicana. The authors describe the housing and husbandry of the colony, the procedures used for mating the animals and the behavior of the animals during mating. They report that the animals showed social behavior and could be bred successfully. Most breeding pairs successfully produced litters; some pairs produced more than one litter. The authors also report data for other parameters, such as the interval between pairing and birth or between births of consecutive litters, litter size, survival to weaning, the timing of sexual maturity and the effects of breeder age on breeding success. PMID:21772350

  13. Convergent life-history shifts: toxic environments result in big babies in two clades of poeciliids

    NASA Astrophysics Data System (ADS)

    Riesch, Rüdiger; Plath, Martin; García de León, Francisco J.; Schlupp, Ingo

    2010-02-01

    The majority of studies on ecological speciation in animals have investigated the divergence caused by biotic factors like divergent food sources or predatory regimes. Here, we examined a system where ecological speciation can clearly be ascribed to abiotic environmental gradients of naturally occurring toxic hydrogen sulfide (H2S). In southern Mexico, two genera of livebearing fishes (Poeciliidae: Poecilia and Gambusia) thrive in various watercourses with different concentrations of H2S. Previous studies have revealed pronounced genetic differentiation between different locally adapted populations in one species ( Poecilia mexicana), pointing towards incipient speciation. In the present study, we examined female reproductive life-history traits in two species pairs: Gambusia sexradiata (from a nonsulfidic and a sulfidic habitat) and Gambusia eurystoma (sulfide-endemic), as well as P. mexicana (nonsulfidic and sulfidic) and Poecilia sulphuraria (sulfide endemic). We found convergent divergence of life-history traits in response to sulfide; most prominently, extremophile poeciliids exhibit drastically increased offspring size coupled with reduced fecundity. Furthermore, within each genus, this trend increased with increasing sulfide concentrations and was most pronounced in the two endemic sulfur-adapted species. We discuss the adaptive significance of large offspring size in toxic environments and propose that divergent life-history evolution may promote further ecological divergence through isolation by adaptation.

  14. An iron-superoxide dismutase antigen-based serological screening of dogs indicates their potential role in the transmission of cutaneous leishmaniasis and trypanosomiasis in Yucatan, Mexico.

    PubMed

    Longoni, Silvia S; Marín, Clotilde; Sauri-Arceo, Carlos H; López-Cespedes, Angeles; Rodríguez-Vivas, Roger I; Villegas, Noelia; Escobedo-Ortegón, Javier; Barrera-Pérez, Mario A; Bolio-Gonzalez, Manuel E; Sánchez-Moreno, Manuel

    2011-07-01

    An increasing number of studies have reported high infection rates for American cutaneous leishmaniasis in dogs, which have thus been proposed as the reservoir host. Canine leishmaniasis is widespread in different states in Mexico, where a number of Leishmania species have been isolated from dogs. In the present study, the detection of different Leishmania species is described in stray dogs from two localities, namely Tulum and Celestún on the Yucatan Peninsula (Mexico). The use of iron-superoxide dismutase excreted by the parasites as the antigen fraction and enzyme-linked immunosorbent assay and western blot tests allowed us to confirm the presence of at least three species of Leishmania (Le. mexicana, Le. braziliensis, and Le. panamensis), some of which are reported for the first time in this species. In addition to a high prevalence of Le. mexicana and Le. braziliensis, and to a lesser degree, Le. panamensis, there is a significant prevalence of Trypanosoma cruzi, suggesting that the dog may be a source of transmission of trypanosomiasis. However, a more thorough epidemiological study on the dog population, both wild as well as urban, of the Yucatan Peninsula will be required to design a control strategy for these diseases, paying particular attention to the population affected and even broadening the study to other Mexican states as well as neighboring countries. These results again confirm that iron-superoxide dismutase excreted by the different trypanosomatid species constitutes a good source of antigen for serodiagnosis in epidemiological studies. PMID:21323424

  15. Phylogenetic analyses of the subgenus Mollienesia (Poecilia, Poeciliidae, Teleostei) reveal taxonomic inconsistencies, cryptic biodiversity, and spatio-temporal aspects of diversification in Middle America.

    PubMed

    Palacios, Maura; Voelker, Gary; Arias Rodriguez, Lenin; Mateos, Mariana; Tobler, Michael

    2016-10-01

    The subgenus Mollienesia is a diverse group of freshwater fishes, including species that have served as important models across multiple biological disciplines. Nonetheless, the taxonomic history of this group has been conflictive and convoluted, in part because the evolutionary relationships have not been rigorously resolved. We conducted a comprehensive molecular phylogenetic analysis of the subgenus Mollienesia to identify taxonomic discrepancies and potentially identify undescribed species, estimate ancestral areas of origin and estimate dates of divergence, as well as explore biogeographical patterns. Our findings confirm the presence of three main clades composed of the P. latipinna, P. sphenops, and P. mexicana species complexes. Unlike previously hypothesized morphology-based analyses, species found on the Caribbean Islands are not part of Mollienesia, but are more closely related to species of the subgenus Limia. Our study also revealed several taxonomic inconsistencies and distinct lineages in the P. mexicana species complex that may represent undescribed species. The diversity in the subgenus Mollienesia is a result of dynamic geologic activity leading to vicariant events, dispersal across geologic blocks, and ecological speciation. PMID:27472959

  16. Genomic variation in recently collected maize landraces from Mexico

    PubMed Central

    Arteaga, María Clara; Moreno-Letelier, Alejandra; Mastretta-Yanes, Alicia; Vázquez-Lobo, Alejandra; Breña-Ochoa, Alejandra; Moreno-Estrada, Andrés; Eguiarte, Luis E.; Piñero, Daniel

    2015-01-01

    The present dataset comprises 36,931 SNPs genotyped in 46 maize landraces native to Mexico as well as the teosinte subspecies Zea maiz ssp. parviglumis and ssp. mexicana. These landraces were collected directly from farmers mostly between 2006 and 2010. We accompany these data with a short description of the variation within each landrace, as well as maps, principal component analyses and neighbor joining trees showing the distribution of the genetic diversity relative to landrace, geographical features and maize biogeography. High levels of genetic variation were detected for the maize landraces (HE = 0.234 to 0.318 (mean 0.311), while slightly lower levels were detected in Zea m. mexicana and Zea m. parviglumis (HE = 0.262 and 0.234, respectively). The distribution of genetic variation was better explained by environmental variables given by the interaction of altitude and latitude than by landrace identity. This dataset is a follow up product of the Global Native Maize Project, an initiative to update the data on Mexican maize landraces and their wild relatives, and to generate information that is necessary for implementing the Mexican Biosafety Law. PMID:26981357

  17. Genome of Leptomonas pyrrhocoris: a high-quality reference for monoxenous trypanosomatids and new insights into evolution of Leishmania

    PubMed Central

    Flegontov, Pavel; Butenko, Anzhelika; Firsov, Sergei; Kraeva, Natalya; Eliáš, Marek; Field, Mark C.; Filatov, Dmitry; Flegontova, Olga; Gerasimov, Evgeny S.; Hlaváčová, Jana; Ishemgulova, Aygul; Jackson, Andrew P.; Kelly, Steve; Kostygov, Alexei Y.; Logacheva, Maria D.; Maslov, Dmitri A.; Opperdoes, Fred R.; O’Reilly, Amanda; Sádlová, Jovana; Ševčíková, Tereza; Venkatesh, Divya; Vlček, Čestmír; Volf, Petr; Jan Votýpka; Záhonová, Kristína; Yurchenko, Vyacheslav; Lukeš, Julius

    2016-01-01

    Many high-quality genomes are available for dixenous (two hosts) trypanosomatid species of the genera Trypanosoma, Leishmania, and Phytomonas, but only fragmentary information is available for monoxenous (single-host) trypanosomatids. In trypanosomatids, monoxeny is ancestral to dixeny, thus it is anticipated that the genome sequences of the key monoxenous parasites will be instrumental for both understanding the origin of parasitism and the evolution of dixeny. Here, we present a high-quality genome for Leptomonas pyrrhocoris, which is closely related to the dixenous genus Leishmania. The L. pyrrhocoris genome (30.4 Mbp in 60 scaffolds) encodes 10,148 genes. Using the L. pyrrhocoris genome, we pinpointed genes gained in Leishmania. Among those genes, 20 genes with unknown function had expression patterns in the Leishmania mexicana life cycle suggesting their involvement in virulence. By combining differential expression data for L. mexicana, L. major and Leptomonas seymouri, we have identified several additional proteins potentially involved in virulence, including SpoU methylase and U3 small nucleolar ribonucleoprotein IMP3. The population genetics of L. pyrrhocoris was also addressed by sequencing thirteen strains of different geographic origin, allowing the identification of 1,318 genes under positive selection. This set of genes was significantly enriched in components of the cytoskeleton and the flagellum. PMID:27021793

  18. The genome sequence of Leishmania (Leishmania) amazonensis: functional annotation and extended analysis of gene models.

    PubMed

    Real, Fernando; Vidal, Ramon Oliveira; Carazzolle, Marcelo Falsarella; Mondego, Jorge Maurício Costa; Costa, Gustavo Gilson Lacerda; Herai, Roberto Hirochi; Würtele, Martin; de Carvalho, Lucas Miguel; Carmona e Ferreira, Renata; Mortara, Renato Arruda; Barbiéri, Clara Lucia; Mieczkowski, Piotr; da Silveira, José Franco; Briones, Marcelo Ribeiro da Silva; Pereira, Gonçalo Amarante Guimarães; Bahia, Diana

    2013-12-01

    We present the sequencing and annotation of the Leishmania (Leishmania) amazonensis genome, an etiological agent of human cutaneous leishmaniasis in the Amazon region of Brazil. L. (L.) amazonensis shares features with Leishmania (L.) mexicana but also exhibits unique characteristics regarding geographical distribution and clinical manifestations of cutaneous lesions (e.g. borderline disseminated cutaneous leishmaniasis). Predicted genes were scored for orthologous gene families and conserved domains in comparison with other human pathogenic Leishmania spp. Carboxypeptidase, aminotransferase, and 3'-nucleotidase genes and ATPase, thioredoxin, and chaperone-related domains were represented more abundantly in L. (L.) amazonensis and L. (L.) mexicana species. Phylogenetic analysis revealed that these two species share groups of amastin surface proteins unique to the genus that could be related to specific features of disease outcomes and host cell interactions. Additionally, we describe a hypothetical hybrid interactome of potentially secreted L. (L.) amazonensis proteins and host proteins under the assumption that parasite factors mimic their mammalian counterparts. The model predicts an interaction between an L. (L.) amazonensis heat-shock protein and mammalian Toll-like receptor 9, which is implicated in important immune responses such as cytokine and nitric oxide production. The analysis presented here represents valuable information for future studies of leishmaniasis pathogenicity and treatment. PMID:23857904

  19. Hepatic hemorrhage, hemocoelom, and sudden death due to Haemoproteus infection in passerine birds: eleven cases.

    PubMed

    Donovan, Taryn A; Schrenzel, Mark; Tucker, Tammy A; Pessier, Allan P; Stalis, Ilse H

    2008-05-01

    Haemoproteus spp. are ancient apicomplexan hemoparasites that have undergone extensive coevolution with their natural hosts and are typically species specific, with inapparent or minimal pathogenicity. A promiscuous genotype of Haemoproteus capable of undergoing host switching on a familial level was identified. This protozoan caused severe disease with high mortality in 6 species of exotic passerine birds housed in California at the San Diego Zoo's Wild Animal Park: Surinam crested oropendola (Psarocolius decumanus decumanus), Guianan turquoise tanager (Tangara mexicana mexicana), blue-necked tanager (Tangara cyanicollis caeruleocephala, Guianan red-capped cardinal (Paroaria gularis gularis), magnificent bird of paradise (Diphyllodes magnificus hunsteini), and superb bird of paradise (Lophorina superba). The birds had few or no clinical signs. Necropsy findings consisted of hemocoelom and irregularly scattered areas of hemorrhage and hepatocellular necrosis. Affected areas of liver contained solitary protozoal megaloschizonts in varied states of degeneration and peripheral nonsuppurative inflammation. No other parasite life stages were found in parenchymal organs or blood smears. Polymerase chain reaction using consensus primers for an avian malarial mitochondrial cytochrome B gene segment was positive in all cases. Sequencing and BLAST analysis identified the protozoan as a Haemoproteus sp. related to Haemoproteus spp. found in asymptomatic passerine birds native to North America. In situ hybridization was performed in 3 animals with a mitochondrial cytochrome B probe and was positive only in megaloschizonts. These findings suggest the recognition of a genotype of Haemoproteus that exhibits high levels of host infidelity and causes severe disease in captive birds exotic to North America. PMID:18460616

  20. Feeding outside the forest: the importance of crop raiding and an invasive weed in the diet of gallery forest ring-tailed lemurs (Lemur catta) following a cyclone at the Beza Mahafaly special reserve, Madagascar.

    PubMed

    LaFleur, M; Gould, L

    2009-01-01

    In January 2005, a cyclone hit southern Madagascar, including the Beza Mahafaly Special Reserve, disrupting the flowering/fruiting cycle of Tamarindus indica, leaving Lemur catta without its major food resource during reproductive periods. We studied two adjacent groups of L. catta during the late gestation period, and both groups ventured outside the reserve to feed. The Red group (RG) fed daily on cultivated sweet potato (Ipomoea batatas) leaves in a nearby field, and both groups consumed leaves and stems of the invasive terrestrial flowering herb Mexican prickly poppy (Argemone mexicana), growing outside the reserve. The Green group (GG) spent significantly more time feeding than did RG, and more time feeding inside the forest compared to outside. The members of RG spent half of their time feeding in the crops, and nearly half of their diet consisted of easy-to-process sweet potato leaves. Additionally, RG defended and restricted GG's access to the crop territory. Of the two non-forest foods, A. mexicana leaves were higher in protein and most minerals (P, Mg, K and Na, but not Ca) and lower in fiber than sweet potato leaves, but sweet potato leaves were preferred by RG. L. catta is a markedly flexible primate with respect to diet, and switches to fallback foods from outside the forest during periods of low food availability. In the highly seasonal and unpredictable climate of southern Madagascar, such behavioral adaptations are important to the survival of this species. PMID:19776607

  1. Genome of Leptomonas pyrrhocoris: a high-quality reference for monoxenous trypanosomatids and new insights into evolution of Leishmania.

    PubMed

    Flegontov, Pavel; Butenko, Anzhelika; Firsov, Sergei; Kraeva, Natalya; Eliáš, Marek; Field, Mark C; Filatov, Dmitry; Flegontova, Olga; Gerasimov, Evgeny S; Hlaváčová, Jana; Ishemgulova, Aygul; Jackson, Andrew P; Kelly, Steve; Kostygov, Alexei Y; Logacheva, Maria D; Maslov, Dmitri A; Opperdoes, Fred R; O'Reilly, Amanda; Sádlová, Jovana; Ševčíková, Tereza; Venkatesh, Divya; Vlček, Čestmír; Volf, Petr; Jan Votýpka; Záhonová, Kristína; Yurchenko, Vyacheslav; Lukeš, Julius

    2016-01-01

    Many high-quality genomes are available for dixenous (two hosts) trypanosomatid species of the genera Trypanosoma, Leishmania, and Phytomonas, but only fragmentary information is available for monoxenous (single-host) trypanosomatids. In trypanosomatids, monoxeny is ancestral to dixeny, thus it is anticipated that the genome sequences of the key monoxenous parasites will be instrumental for both understanding the origin of parasitism and the evolution of dixeny. Here, we present a high-quality genome for Leptomonas pyrrhocoris, which is closely related to the dixenous genus Leishmania. The L. pyrrhocoris genome (30.4 Mbp in 60 scaffolds) encodes 10,148 genes. Using the L. pyrrhocoris genome, we pinpointed genes gained in Leishmania. Among those genes, 20 genes with unknown function had expression patterns in the Leishmania mexicana life cycle suggesting their involvement in virulence. By combining differential expression data for L. mexicana, L. major and Leptomonas seymouri, we have identified several additional proteins potentially involved in virulence, including SpoU methylase and U3 small nucleolar ribonucleoprotein IMP3. The population genetics of L. pyrrhocoris was also addressed by sequencing thirteen strains of different geographic origin, allowing the identification of 1,318 genes under positive selection. This set of genes was significantly enriched in components of the cytoskeleton and the flagellum. PMID:27021793

  2. Skeletal morphology of the forefoot in shrews (Mammalia: Soricidae) of the genus Cryptotis, as revealed by digital x-rays

    USGS Publications Warehouse

    Woodman, N.; Morgan, J.P.J.

    2005-01-01

    Variation in the forefoot skeleton of small-eared shrews (family Soricidae, genus Cryptotis) has been previously documented, but the paucity of available skeletons for most taxa makes assessment of the degrees of intraspecific and interspecific variation difficult. We used a digital X-ray system to extract images of the forefoot skeleton from 101 dried skins of eight taxa (seven species, including two subspecies of one species) of these shrews. Lengths and widths of each of the four bones of digit III were measured directly from the digital images, and we used these data to quantify variation within and among taxa. Analysis of the images and measurements showed that interspecific variation exceeds intraspecific variation. In fact, most taxa could be distinguished in multivariate and some bivariate plots. Our quantitative data helped us define a number of specific forefoot characters that we subsequently used to hypothesize evolutionary relationships among the taxa using the exhaustive search option in PAUP, a computer program for phylogenetic analysis. The resulting trees generally concur with previously published evolutionary hypotheses for small-eared shrews. Cryptotis meridensis, a taxon not previously examined in recent phylogenies, is rooted at the base of the branch leading to the C. mexicana group of species. The position of this species suggests that the mostly South American C. thomasi group shares an early ancestor with the C. mexicana group.

  3. Biological aspects of crosses between Triatoma recurva (Stål), 1868 (Hemiptera: Reduviidae: Triatominae) and other members of the Phyllosoma complex.

    PubMed

    Martínez-Ibarra, José Alejandro; Nogueda-Torres, Benjamín; Licón-Trillo, Ángel; Alejandre-Aguilar, Ricardo; Salazar-Schettino, Paz María; Vences-Blanco, Mauro Omar

    2015-06-01

    The degree of reproductive isolation between Triatoma recurva (Stål) (Hemiptera: Reduviidae: Triatominae) and the six species of the genus Meccus plus T. mexicana (Herrich-Schaeffer) (Hemiptera: Reduviidae) was examined. Fertility and the segregation of morphological characteristics were examined in two generations of hybrids from crosses between these species. The percentage of couples with offspring (fertile) was low in the vast majority of sets of crosses, with the exception of that between T. recurva female and M. phyllosomus male. In all studied sets of crosses, no first- (F1) or second- (F2) generation individuals were morphologically similar to T. recurva but instead shared the morphology of the other parental species. A similar phenomenon was observed in the three successful sets of backcrosses. These results indicated that different recorded levels of reproductive fitness among T. recurva and the species of Meccus involved in this study, plus T. mexicana, are present and that they were apparently influenced by differing mechanisms of isolation. The presence of some degree of reproductive compatibility between studied triatomines of distinct genera (Meccus spp. and Triatoma spp.) reinforces the need for generic revision of the tribe Triatomini. PMID:26047191

  4. Identification of Leishmania spp. promastigotes in the intestines, ovaries and salivary glands of Rhipicephalus sanguineus actively infesting dogs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sand flies are recognized as the major vector of canine visceral leishmaniasis. However, in some areas of Brazil where sand flies do not occur, this disease is found in humans and dogs. There has been speculation that ticks might play a role in transmission of canine visceral leishmaniasis and the D...

  5. Drug Resistance in Natural Isolates of Leishmania donovani s.l. Promastigotes Is Dependent of Pgp170 Expression

    PubMed Central

    Mazeris, Apostolos; Koutala, Eleni; Vlahou, Antonia; Papadogiorgaki, Sevasti; Antoniou, Maria

    2013-01-01

    Resistance of pathogens to drugs is a growing concern regarding many diseases. Parasites like Leishmania, Plasmodium and Entamoeba histolytica; and neoplastic cells, present the multidrug-resistant phenotype rendering chemotherapy ineffective. The acquired resistance of Leishmania to antimony has generated intense research on the mechanisms involved but the question has not yet been resolved. To test the hypothesis that drug efflux in Leishmania, as measured by flow cytometry using the fluorescent dye Rhodamine-123, is largely dependent on the number of efflux pumps an isolate can express, the amount of Pgp 170 molecules was assessed in ten field isolates (5 “resistant” and 5 “susceptible”) using: Western Blotting, Confocal and Transmission Electron Microscopy, and proteomics. Their survival after exposure to three antileishmanial drugs, in vitro, was evaluated and clinical data were compared to the in vitro results. All isolates were resistant to Glucantime but susceptible to Miltefosine, whilst Amphotericin B was more effective on the “susceptible” isolates. The MDR gene, expressing the transmembrane efflux pump Pgp 170, appears to play a key role in the phenomenon of drug resistance. When “susceptible” versus “resistant” parasites were compared, it was shown that the higher the number of Pgp 170 molecules the higher the Rhodamine-123 efflux from the parasite body and, when exposed to the drug, the number of efflux pumps increased. However, the rate of this increase was not linear and it is possible that there is a maximum number of Pgp 170 molecules an isolate can express. Nevertheless, the phenomenon is a complex one and other factors and proteins are involved in which the HSP-70 group proteins, detected in the “resistant” isolates, may play a significant role. PMID:23776486

  6. Leishmanial Excreted Factor: Protein-Bound and Free Forms from Promastigote Cultures of Leishmania tropica and Leishmania donovani

    PubMed Central

    Slutzky, Gerald M.; El-On, Joseph; Greenblatt, Charles L.

    1979-01-01

    Leishmania spp. growing in culture produce an immunologically active substance called excreted factor (EF), which precipitates antibodies raised against intact cells and has been implicated as the conditioning agent for parasite infection of host macrophages. An improved method for isolation of the material is described, based on Sephadex column chromatography of growth medium which had been boiled at pH 5.0. This procedure allows the detection of differences among the EF molecules of different species, and it overcomes previous shortcomings through the monitoring of immunological activity throughout. Analysis of the products of this procedure revealed that EFs from Leishmania tropica and Leishmania donovani share a common carrier protein, identified as rabbit serum albumin, and are chemically quite similar. Growth medium from L. tropica boiled at acidic pH contains primarily an EF-albumin complex of 75,000 molecular weight. Treated growth medium from L. donovani, on the other hand, contains both the albumin complex and a smaller molecule (less than 27,000 molecular weight) that is not associated with rabbit protein. This material accounts for nearly 20% of the EF of one L. donovani strain, but constitutes only a minute fraction of L. tropica EF. Treatment of the EF-albumin complex with trichloroacetic acid separates the molecule into two major subunits, one having a molecular weight of about 61,000 (without anti-Leishmania activity) and the other having a molecular weight of about 18,000 (with no anti-rabbit activity). The protein-free EF of L. tropica differs from that released by trichloroacetic acid extraction in that it is capable of precipitating antisera of nonhomologous serotypes, whereas the albumin complex and the trichloroacetic acid-treated EF fragment are not. EFs from both species display pH-dependent affinity for certain lectins. Images PMID:118936

  7. CD8 Cells of Patients with Diffuse Cutaneous Leishmaniasis Display Functional Exhaustion: The Latter Is Reversed, In Vitro, by TLR2 Agonists

    PubMed Central

    Hernández-Ruiz, Joselín; Salaiza-Suazo, Norma; Carrada, Georgina; Escoto, Sofía; Ruiz-Remigio, Adriana; Rosenstein, Yvonne; Zentella, Alejandro; Becker, Ingeborg

    2010-01-01

    Leishmania mexicana (Lm) causes localized (LCL) and diffuse (DCL) cutaneous leishmaniasis. DCL patients have a poor cellular immune response leading to chronicity. It has been proposed that CD8 T lymphocytes (CD8) play a crucial role in infection clearance, although the role of CD8 cytotoxicity in disease control has not been elucidated. Lesions of DCL patients have been shown to harbor low numbers of CD8, as compared to patients with LCL, and leishmanicidal treatment restores CD8 numbers. The marked response of CD8 towards Leishmania parasites led us to analyze possible functional differences between CD8 from patients with LCL and DCL. We compared IFNγ production, antigen-specific proliferation, and cytotoxicity of CD8 purified from PBMC against autologous macrophages (MO) infected with Leishmania mexicana (MOi). Additionally, we analyzed tissue biopsies from both groups of patients for evidence of cytotoxicity associated with apoptotic cells in the lesions. We found that CD8 cell of DCL patients exhibited low cytotoxicity, low antigen-specific proliferation and low IFNγ production when stimulated with MOi, as compared to LCL patients. Additionally, DCL patients had significantly less TUNEL+ cells in their lesions. These characteristics are similar to cellular “exhaustion” described in chronic infections. We intended to restore the functional capacity of CD8 cells of DCL patients by preincubating them with TLR2 agonists: Lm lipophosphoglycan (LPG) or Pam3Cys. Cytotoxicity against MOi, antigen-specific proliferation and IFNγ production were restored with both stimuli, whereas PD-1 (a molecule associated with cellular exhaustion) expression, was reduced. Our work suggests that CD8 response is associated with control of Lm infection in LCL patients and that chronic infection in DCL patients leads to a state of CD8 functional exhaustion, which could facilitate disease spread. This is the first report that shows the presence of functionally exhausted CD8 T

  8. Review and redescription of species in the Oecetis avara group, with the description of 15 new species (Trichoptera, Leptoceridae)

    PubMed Central

    Blahnik, Roger J.; Holzenthal, Ralph W.

    2014-01-01

    Abstract The O. avara group of Oecetis is formally defined to include 4 described species, O. avara (Banks), O. disjuncta (Banks), O. elata Denning & Sykora, and O. metlacenis Bueno-Soria, and 15 new species. Oecetis marquesi Bueno-Soria, previously considered a member of the O. avara group, is treated as incertae sedis to species group, but is also redescribed and treated in the current work. New species described here (with their respective distributions) include: O. acciptrina (Costa Rica, Panama, Ecuador), O. agosta (Mexico), O. angularis (Guatemala to Ecuador), O. apache (SW USA), O. campana (Ecuador), O. constricta (Mexico to Ecuador, Venezuela, and Trinidad), O. houghtoni (North America), O. maritza (Costa Rica), O. mexicana (Mexico to Ecuador), O. patula (Guatemala, Nicaragua), O. protrusa (Mexico to Ecuador), O. sordida (Mexico, USA, Canada), O. tumida (Costa Rica), O. uncata (Costa Rica), and O. verrucula (Mexico to Costa Rica). A key to the species is also provided. PMID:24574849

  9. Pattern Recognition Techniques Applied to the Study of Leishmanial Glyceraldehyde-3-Phosphate Dehydrogenase Inhibition

    PubMed Central

    Lozano, Norka B. H.; Oliveira, Rafael F.; Weber, Karen C.; Honorio, Kathia M.; Guido, Rafael V. C.; Andricopulo, Adriano D.; de Sousa, Alexsandro G.; da Silva, Albérico B. F.

    2014-01-01

    Chemometric pattern recognition techniques were employed in order to obtain Structure-Activity Relationship (SAR) models relating the structures of a series of adenosine compounds to the affinity for glyceraldehyde 3-phosphate dehydrogenase of Leishmania mexicana (LmGAPDH). A training set of 49 compounds was used to build the models and the best ones were obtained with one geometrical and four electronic descriptors. Classification models were externally validated by predictions for a test set of 14 compounds not used in the model building process. Results of good quality were obtained, as verified by the correct classifications achieved. Moreover, the results are in good agreement with previous SAR studies on these molecules, to such an extent that we can suggest that these findings may help in further investigations on ligands of LmGAPDH capable of improving treatment of leishmaniasis. PMID:24566143

  10. Systematic position of Mucronella perforata Okada et Mawatari 1937

    NASA Astrophysics Data System (ADS)

    Liu, Xi-Xing; Liu, Hui-Lian

    1999-12-01

    Mucronella perforata was first discovered from Onagawa Bay, northern Honshu, Japan, in 1937 by Okada and Mawatari. It was reported from Hong Kong and its neighboring waters and was referred to the genus Hippopodina Levinsen, 1909 of the Family Hippopodinidae Levinsen, 1909 by Huang et al. (1990) and Huang (1994). According to the astogeny of the species, both the ancestrula and its successor zooids have an ascus proximal to the orifice, and its hyperstomial ovicells are imperforate, so it could not be a member of any known genus of the known families of the superfamily Schizoporelloidea Jullien, 1883 in the infraorder Lepraliomorpha Gordon, 1989. The new genus Pacificincola and the new family Pacificincolidae established respectively on the basis of reexaminations of the Chinese specimens obtained from the southern Yellow Sea, the East China Sea and the northern South China Sea. Schizoporella insculpta Hincks, 1880 and Hippoporina mexicana Soule, Soule et Chaney, 1995 have been referred to the new genus in the paper.

  11. Mexican plants with hypoglycaemic effect used in the treatment of diabetes.

    PubMed

    Andrade-Cetto, Adolfo; Heinrich, Michael

    2005-07-14

    Diabetes mellitus is a syndrome which affects more and more people in all countries over the world. In México, it is commonly treated with herbal extracts. Such treatment may be of considerable benefit especially during the early stages of the illness. In this review, we discuss species commonly used in México in the treatment of diabetes. A total of 306 species have records of a popular use in the treatment of this syndrome in México. Seven of these species--Cecropia obtusifolia Bertol. (Cecropiaceae), Equisetum myriochaetum Schlecht & Cham (Equisetaceae), Acosmium panamense (Benth.) Yacolev (Fabaceae), Cucurbita ficifolia Bouché (Cucurbitaceae), Agarista mexicana (Hemsl.) Judd. (Ericaeae), Brickellia veronicaefolia (Kunth) A. Gray (Asteraceae), Parmentiera aculeata (Kunth) Seem. (Bignoniaceae)--are discussed in greater detail, highlighting our current knowledge about these botanicals, but also the enormous gaps in our knowledge, most notably as it relates to the species' toxicology, the pharmacokinetics of its active constituents and their metabolism. PMID:15964161

  12. Assessing the competitive ability of Japanese stilt grass, Microstegium vimineum (Trin.) A. Camus

    USGS Publications Warehouse

    Leicht, S.A.; Silander, J.A., Jr.; Greenwood, K.

    2005-01-01

    Microstegium vimineum (Japanese stilt grass) is an invasive grass in the eastern half of the United States which can form dense monocultures in forest understories, displacing native species. Although the loss of native species has been observed in the field, the actual competitive ability of this grass has not been examined. Microstegium vimineum was grown under controlled environment, greenhouse conditions in competition with Lolium perenne ssp. multiflorum (annual rye grass) and Muhlenbergia mexicana (Mexican muhly) in varying density ratios in full and low light treatments. Microstegium vimineum had a greater aboveground biomass, relative growth rate, and reproductive output than both competitors in both light treatments. The high competitive ability of Microstegium vimineum, especially in low light conditions, reflects its highly aggressive nature in forested or other landscapes of eastern North America.

  13. A survey of selected parasitic and viral pathogens in four species of Mexican parrots, Amazona autumnalis, Amazona oratrix, Amazona viridigenalis, and Rhynchopsitta pachyrhyncha.

    PubMed

    Stone, Elizabeth Gordon; Montiel-Parra, Griselda; Pérez, Tila Maria

    2005-06-01

    Isolated populations of four species of Mexican parrots were sampled for evidence of selected pathogens of concern in birds originating in Latin America. Data were collected between June and September 1997, and ectoparasite collection was repeated with Rhynchopsitta pachyrhyncha in September 2000. Serum samples from nine Amazona oratrix, 10 Amazona viridigenalis, 6 Amazona autumnalis, and 25 R. pachyrhyncha chicks were screened for neutralizing antibodies to psittacid herpesvirus and avian influenza and for antibodies to paramyxovirus serotypes 1 and 3. Chicks were also examined visually for fecal parasites and ectoparasites. All serologic and fecal parasite tests were negative. Ectoparasites included ticks, Ixodidae; mites, Ornithonyssus sylviarum; fleas, Psyttopsylla mexicana; lice, Paragoniocotes mexicanus, Heteromenopon sp., and Psittacobrosus sp.; and bugs, Ornithocoris sp. This study provides baseline information to guide future health studies. PMID:17323565

  14. A New Antileishmanial Preparation of Combined Solamargine and Solasonine Heals Cutaneous Leishmaniasis through Different Immunochemical Pathways.

    PubMed

    Lezama-Dávila, C M; McChesney, J D; Bastos, J K; Miranda, M A; Tiossi, R F; da Costa, J de C; Bentley, M V; Gaitan-Puch, S E; Isaac-Márquez, A P

    2016-05-01

    Little has been done during the past 100 years to develop new antileishmanial drugs. Most infected individuals live in poor countries and have a low cash income to be attractive targets to pharmaceutical corporations. Two heterosidic steroids, solamargine and solasonine, initially identified as major components of the Brazilian plant Solanum lycocarpum, were tested for leishmanicidal activity. Both alkaloids killed intracellular and extracellular Leishmania mexicana parasites more efficiently than the reference drug sodium stibogluconate. A total of 10 μM each individual alkaloid significantly reduced parasite counts in infected macrophages and dendritic cells. In vivo treatment of C57BL/6 mice with a standardized topical preparation containing solamargine (45.1%) and solasonine (44.4%) gave significant reductions in lesion sizes and parasite counts recovered from lesions. Alkaloids present different immunochemical pathways in macrophages and dendritic cells. We conclude that this topical preparation is effective and a potential new and inexpensive treatment for cutaneous leishmaniasis. PMID:26883711

  15. Molecular Components of the Sporothrix schenckii Complex that Induce Immune Response.

    PubMed

    Alba-Fierro, Carlos A; Pérez-Torres, Armando; Toriello, Conchita; Romo-Lozano, Yolanda; López-Romero, Everardo; Ruiz-Baca, Estela

    2016-08-01

    Sporotrichosis is a fungal disease caused by the Sporothrix schenckii complex that includes species such as S. brasiliensis, S. schenckii sensu stricto, S. globosa, S. luriei, S. mexicana, and S. pallida, which exhibit different potentially antigenic molecular components. The immune response of susceptible hosts to control infection and disease caused by these fungi has been little studied. Besides, the fungus-host interaction induces the activation of different types of immune response. This mini-review analyzes and discusses existing reports on the identification and functional characterization of molecules from species of the S. schenckii complex with clinical relevance, and the mechanisms that mediate the type and magnitude of the immune response in experimental models in vivo and in vitro. This knowledge is expected to contribute to the development of protective and therapeutic strategies against sporotrichosis and other mycoses. PMID:27117164

  16. Divergent evolution of male aggressive behaviour: another reproductive isolation barrier in extremophile poeciliid fishes?

    PubMed

    Bierbach, David; Klein, Moritz; Saßmannshausen, Vanessa; Schlupp, Ingo; Riesch, Rüdiger; Parzefall, Jakob; Plath, Martin

    2012-01-01

    Reproductive isolation among locally adapted populations may arise when immigrants from foreign habitats are selected against via natural or (inter-)sexual selection (female mate choice). We asked whether also intrasexual selection through male-male competition could promote reproductive isolation among populations of poeciliid fishes that are locally adapted to extreme environmental conditions [i.e., darkness in caves and/or toxic hydrogen sulphide (H(2)S)]. We found strongly reduced aggressiveness in extremophile P. oecilia mexicana, and darkness was the best predictor for the evolutionary reduction of aggressiveness, especially when combined with presence of H(2)S. We demonstrate that reduced aggression directly translates into migrant males being inferior when paired with males from non-sulphidic surface habitats. By contrast, the phylogenetically old sulphur endemic P. sulphuraria from another sulphide spring area showed no overall reduced aggressiveness, possibly indicating evolved mechanisms to better cope with H(2)S. PMID:22315695

  17. Molecular identification and barcodes for the genus Nymphaea.

    PubMed

    Chaveerach, Arunrat; Tanee, T; Sudmoon, Runglawan

    2011-09-01

    Nymphaea species, the most popular decorative plants, were collected for specificity of inter-simple sequence repeat (ISSR) analyses in species identification and differentiation of cultivars and natural populations. Dendrogram constructed from ISSR analyses separated out wild species, namely Nymphaea cyanea, N. nouchali, N. capensis, N. lotus and an outgroup N. mexicana, and cultivars. The dendrogram indicates that the cultivars should be differentiated from N. capensis, as they are sister individuals of N. capensis. The ISSR banding data and the dendrogram are concordantly concluded that wild N. capensis would be an effective type species for producing different cultivars. After plant identification by ISSR markers, DNA barcodes of all sample materials were done to provide species specific markers which can be used for rapid and accurate further plant identification without morphological characters. DNA barcoding sequence analysis indicates genetic distance values. All sequences were recorded in GenBank database. PMID:21840834

  18. Anti-Trypanosoma cruzi activity of 10 medicinal plants used in northeast Mexico.

    PubMed

    Molina-Garza, Zinnia Judith; Bazaldúa-Rodríguez, Aldo Fabio; Quintanilla-Licea, Ramiro; Galaviz-Silva, Lucio

    2014-08-01

    The aim of this study was to screen the trypanocidal activity of plants used in traditional Mexican medicine for the treatment of various diseases related to parasitic infections. Cultured Trypanosoma cruzi epimastigotes were incubated for 96h with different concentrations of methanolic extracts obtained from Artemisia mexicana, Castela texana, Cymbopogon citratus, Eryngium heterophyllum, Haematoxylum brasiletto, Lippia graveolens, Marrubium vulgare, Persea americana, Ruta chalepensis and Schinus molle. The inhibitory concentration (IC50) was determined for each extract via a colorimetric method. Among the evaluated species, the methanolic extracts of E. heterophyllum, H. brasiletto, M. vulgare and S. molle exhibited the highest trypanocidal activity, showing percentages of growth inhibition between 88 and 100% at a concentration of 150μg/ml. These medicinal plants may represent a valuable source of new bioactive compounds for the therapeutic treatment of trypanosomiasis. PMID:24742906

  19. First species of Leptochelia Dana, 1849 (Crustacea: Tanaidacea) from the Eastern Pacific, with an annotated checklist and identification keys for the genus.

    PubMed

    Jarquín-González, Jani; García-Madrigal, María Del Socorro; Carrera-Parra, Luis Fernando

    2015-01-01

    Forty three species of leptocheliids are known worldwide. In the American region only eight species have been described from the Western Atlantic, while for the Eastern Pacific none have been described, suggesting that the diversity of this family has been severely underestimated in this region. Here we describe the first species of Leptochelia from the Eastern Pacific, Leptochelia mexicana n. sp., which is characterized by the males having a spiniform seta on the second segment of uropodal endopod, a novel feature for the genus. In addition, the first annotated checklist and a taxonomic key with illustrations for Leptochelia species are included. The list includes the type locality, type depository, distribution, habitat and, in some cases, remarks. PMID:25781398

  20. Improved traditional phytomedicines in current use for the clinical treatment of malaria.

    PubMed

    Willcox, Merlin

    2011-04-01

    Phytomedicines and "green pharmacies" are promoted by some NGOs and governments as part of their efforts to control malaria. "Improved traditional medicines" (ITMs) are standardised as regards preparation and dose, although not always according to the concentration of active compounds. A systematic literature search revealed that six such phytomedicines are currently government-approved in at least one country and used on a relatively large scale nationally or internationally: Artemisia annua L. (Asteraceae), Cinchona bark (Rubiaceae), Cryptolepis sanguinolenta (Lindl.) Schltr. (Apocynaceae), "Ayush-64", "Malarial-5" and Cochlospermum planchonii Hook. f. ex Planch. (Bixaceae). One further ITM has been developed and is in the process of being approved: Argemone mexicana decoction. Their development, phytochemistry, pharmacology, and clinical trials are reviewed, as well as priorities for future research. PMID:21204042

  1. A new genus and two new species of hymenosomatid crabs (Crustacea:Brachyura: Hymenosomatidae) from the southwestern Atlantic and eastern Australia.

    PubMed

    Tavares, Marcos; Santana, William

    2015-01-01

    A new genus, Teramnonotus n. gen., is erected for Elamena gordonae Monod, 1956. The new genus includes two new species, T. johnlucasi n. gen., n. sp. and T. monodi n. gen., n. sp., described herein from eastern Australia (Queensland) and the southwestern Atlantic (Brazil), respectively. Teramnonotus n. gen. superficially resembles Elamena H. Milne Edwards, 1837, and Trigonoplax H. Milne Edwards, 1853, of which it can be easily distinguished by a combination of characters: eyes and ocular peduncle, rostrum, carapace, thoracic sternite 8 and thoracic pleurite 8, and the inhalant water openings. The validity of the obscure species Elamena mexicana H. Milne Edwards, 1853, is not supported and it is synonymised with Halicarcinus planatus (Fabricius, 1775). PMID:25661023

  2. Emergence of Autochthonous Cutaneous Leishmaniasis in Northeastern Texas and Southeastern Oklahoma

    PubMed Central

    Clarke, Carmen F.; Bradley, Kristy K.; Wright, James H.; Glowicz, Janet

    2013-01-01

    Autochthonous human cases of leishmaniasis in the United States are uncommon. We report three new cases of cutaneous leishmaniasis and details of a previously reported case, all outside the known endemic range in Texas. Surveys for enzootic rodent reservoirs and sand fly vectors were conducted around the residences of three of the case-patients during the summer of 2006; female Lutzomyia anthophora sand flies were collected at a north Texas and southeast Oklahoma residence of a case-patient, indicating proximity of a suitable vector. Urban sprawl, climatologic variability, or natural expansion of Leishmania mexicana are possible explanations for the apparent spread to the north and east. Enhanced awareness among healthcare providers in the south central region of the United States is important to ensure clinical suspicion of leishmaniasis, diagnosis, and appropriate patient management. PMID:23185078

  3. Genetic divergence in populations of Lutzomyia ayacuchensis, a vector of Andean-type cutaneous leishmaniasis, in Ecuador and Peru.

    PubMed

    Kato, Hirotomo; Cáceres, Abraham G; Gomez, Eduardo A; Mimori, Tatsuyuki; Uezato, Hiroshi; Hashiguchi, Yoshihisa

    2015-01-01

    Haplotype and gene network analyses were performed on mitochondrial cytochrome oxidase I and cytochrome b gene sequences of Lutzomyia (Lu.) ayacuchensis populations from Andean areas of Ecuador and southern Peru where the sand fly species transmit Leishmania (Leishmania) mexicana and Leishmania (Viannia) peruviana, respectively, and populations from the northern Peruvian Andes, for which transmission of Leishmania by Lu. ayacuchensis has not been reported. The haplotype analyses showed higher intrapopulation genetic divergence in northern Peruvian Andes populations and less divergence in the southern Peru and Ecuador populations, suggesting that a population bottleneck occurred in the latter populations, but not in former ones. Importantly, both haplotype and phylogenetic analyses showed that populations from Ecuador consisted of clearly distinct clusters from southern Peru, and the two populations were separated from those of northern Peru. PMID:25312337

  4. Herbicides to curb human parasitic infections: in vitro and in vivo effects of trifluralin on the trypanosomatid protozoans.

    PubMed Central

    Chan, M M; Grogl, M; Chen, C C; Bienen, E J; Fong, D

    1993-01-01

    Leishmaniasis is a major tropical disease for which current chemotherapies, pentavalent antimonials, are inadequate and cause severe side effects. It has been reported that trifluralin, a microtubule-disrupting herbicide, is inhibitory to Leishmania amazonensis. In this study, the in vitro effect of trifluralin on different species of trypanosomatid protozoans was determined. In addition to L. amazonensis, trifluralin is effective against Leishmania major and Leishmania tropica, which cause cutaneous infections, Leishmania donovani, which causes visceral disease, Leishmania panamensis, which may cause mucocutaneous infection, and Trypanosoma brucei, an important human and veterinary pathogen. Moreover, most encouragingly, trifluralin is effective in vivo as a topical ointment against L. major and Leishmania mexicana murine cutaneous leishmaniasis. Thus, trifluralin is a promising lead drug for several related, prevalent tropical diseases: leishmaniasis, trypanosomiasis of animals, and, possibly, African trypanosomiasis in humans. Images Fig. 4 PMID:8516314

  5. An ectomycorrhizal nitrogen economy facilitates monodominance in a neotropical forest.

    PubMed

    Corrales, Adriana; Mangan, Scott A; Turner, Benjamin L; Dalling, James W

    2016-04-01

    Tropical forests are renowned for their high diversity, yet in many sites a single tree species accounts for the majority of the individuals in a stand. An explanation for these monodominant forests remains elusive, but may be linked to mycorrhizal symbioses. We tested three hypotheses by which ectomycorrhizas might facilitate the dominance of the tree, Oreomunnea mexicana, in montane tropical forest in Panama. We tested whether access to ectomycorrhizal networks improved growth and survival of seedlings, evaluated whether ectomycorrhizal fungi promote seedling growth via positive plant-soil feedback, and measured whether Oreomunnea reduced inorganic nitrogen availability. We found no evidence that Oreomunnea benefits from ectomycorrhizal networks or plant-soil feedback. However, we found three-fold higher soil nitrate and ammonium concentrations outside than inside Oreomunnea-dominated forest and a correlation between soil nitrate and Oreomunnea abundance in plots. Ectomycorrhizal effects on nitrogen cycling might therefore provide an explanation for the monodominance of ectomycorrhizal tree species worldwide. PMID:26833573

  6. Calmodulin Polymerase Chain Reaction-Restriction Fragment Length Polymorphism for Leishmania Identification and Typing.

    PubMed

    Miranda, Aracelis; Samudio, Franklyn; González, Kadir; Saldaña, Azael; Brandão, Adeilton; Calzada, Jose E

    2016-08-01

    A precise identification of Leishmania species involved in human infections has epidemiological and clinical importance. Herein, we describe a preliminary validation of a restriction fragment length polymorphism assay, based on the calmodulin intergenic spacer region, as a tool for detecting and typing Leishmania species. After calmodulin amplification, the enzyme HaeIII yielded a clear distinction between reference strains of Leishmania mexicana, Leishmania amazonensis, Leishmania infantum, Leishmania lainsoni, and the rest of the Viannia reference species analyzed. The closely related Viannia species: Leishmania braziliensis, Leishmania panamensis, and Leishmania guyanensis, are separated in a subsequent digestion step with different restriction enzymes. We have developed a more accessible molecular protocol for Leishmania identification/typing based on the exploitation of part of the calmodulin gene. This methodology has the potential to become an additional tool for Leishmania species characterization and taxonomy. PMID:27352873

  7. Spasmolytic activity of some plants used by the Otomi Indians of Quéretaro (México) for the treatment of gastrointestinal disorders.

    PubMed

    Rojas, A; Bah, M; Rojas, J I; Serrano, V; Pacheco, S

    1999-11-01

    The present study was performed to investigate the effect of chloroform-methanol (1:1) extracts derived from Baccharis heterophylla H. B. K (Asteraceae), Desmodium grahami Gray (Leguminosae), Dyssodia pinnata var. pinnata Rob. (Asteraceae), Gentiana spathacea H. B. K. (Gentianaceae), Loeselia mexicana Brand. (Polemoniaceae), Selaginella pallescens Spring. (Selaginellaceae) and Zornia diphylla Pers. (Fabaceae) on the spontaneous contractions of isolated rat ileum. The concentration-response curves and IC50 values were obtained and the respective relaxant potencies were compared with that of the CHCl3-MeOH (1:1) extract of Datura lanosa. The data showed that all the extracts produce a concentration-dependent inhibition of spontaneous ileum contractions. In addition, the antimicrobial activity of the extracts against pathogenic enterobacteria was tested. The pharmacological actions shown by the extracts of the selected species tend to support ethnomedical use of the plants for the treatment of gastrointestinal disorders. PMID:11962546

  8. Response of cutaneous leishmaniasis (chiclero's ulcer) to treatment with meglumine antimoniate in Southeast Mexico.

    PubMed

    Vargas-Gonzalez, A; Canto-Lara, S B; Damian-Centeno, A G; Andrade-Narvaez, F J

    1999-12-01

    Cutaneous leishmaniasis, known as chiclero's ulcer in southeastern Mexico, is characterized by a predominantly single, painless, ulcerated lesion, without lymphangitis or adenopathy. When located on the ear, it tends to become chronic, causing destruction of the pinna and disfigurement. It is caused predominantly by Leishmania (L.) mexicana. Although pentavalent antimonials (Sb5+) are the mainstay of leishmanial therapy and have been used for more than 50 years, dosage regimens have been repeatedly modified and the best one has not been fully identified. The main purpose of the present study was to investigate the response of chiclero's ulcer to treatment with meglumine antimoniate. One hundred five patients were treated with meglumine antimoniate at a daily dose of 1 ampule per day (425 mg of Sb5+) until healing. The lesions healed after a mean of 25 days (range = 5-60 days). PMID:10674678

  9. Glucose-6-Phosphate Dehydrogenase of Trypanosomatids: Characterization, Target Validation, and Drug Discovery

    PubMed Central

    Gupta, Shreedhara; Igoillo-Esteve, Mariana; Michels, Paul A. M.; Cordeiro, Artur T.

    2011-01-01

    In trypanosomatids, glucose-6-phosphate dehydrogenase (G6PDH), the first enzyme of the pentosephosphate pathway, is essential for the defense of the parasite against oxidative stress. Trypanosoma brucei, Trypanosoma cruzi, and Leishmania mexicana G6PDHs have been characterized. The parasites' G6PDHs contain a unique 37 amino acid long N-terminal extension that in T. cruzi seems to regulate the enzyme activity in a redox-state-dependent manner. T. brucei and T. cruzi G6PDHs, but not their Leishmania spp. counterpart, are inhibited, in an uncompetitive way, by steroids such as dehydroepiandrosterone and derivatives. The Trypanosoma enzymes are more susceptible to inhibition by these compounds than the human G6PDH. The steroids also effectively kill cultured trypanosomes but not Leishmania and are presently considered as promising leads for the development of new parasite-selective chemotherapeutic agents. PMID:22091394

  10. Case report: Emergence of autochthonous cutaneous leishmaniasis in northeastern Texas and southeastern Oklahoma.

    PubMed

    Clarke, Carmen F; Bradley, Kristy K; Wright, James H; Glowicz, Janet

    2013-01-01

    Autochthonous human cases of leishmaniasis in the United States are uncommon. We report three new cases of cutaneous leishmaniasis and details of a previously reported case, all outside the known endemic range in Texas. Surveys for enzootic rodent reservoirs and sand fly vectors were conducted around the residences of three of the case-patients during the summer of 2006; female Lutzomyia anthophora sand flies were collected at a north Texas and southeast Oklahoma residence of a case-patient, indicating proximity of a suitable vector. Urban sprawl, climatologic variability, or natural expansion of Leishmania mexicana are possible explanations for the apparent spread to the north and east. Enhanced awareness among healthcare providers in the south central region of the United States is important to ensure clinical suspicion of leishmaniasis, diagnosis, and appropriate patient management. PMID:23185078

  11. Targeting the Cytochrome bc1 Complex of Leishmania Parasites for Discovery of Novel Drugs.

    PubMed

    Ortiz, Diana; Forquer, Isaac; Boitz, Jan; Soysa, Radika; Elya, Carolyn; Fulwiler, Audrey; Nilsen, Aaron; Polley, Tamsen; Riscoe, Michael K; Ullman, Buddy; Landfear, Scott M

    2016-08-01

    Endochin-like quinolones (ELQs) are potent and specific inhibitors of cytochrome bc1 from Plasmodium falciparum and Toxoplasma gondii and show promise for novel antiparasitic drug development. To determine whether the mitochondrial electron transport chain of Leishmania parasites could be targeted similarly for drug development, we investigated the activity of 134 structurally diverse ELQs. A cohort of ELQs was selectively toxic to amastigotes of Leishmania mexicana and L. donovani, with 50% inhibitory concentrations (IC50s) in the low micromolar range, but the structurally similar hydroxynaphthoquinone buparvaquone was by far the most potent inhibitor of electron transport, ATP production, and intracellular amastigote growth. Cytochrome bc1 is thus a promising target for novel antileishmanial drugs, and further improvements on the buparvaquone scaffold are warranted for development of enhanced therapeutics. PMID:27297476

  12. Review and redescription of species in the Oecetis avara group, with the description of 15 new species (Trichoptera, Leptoceridae).

    PubMed

    Blahnik, Roger J; Holzenthal, Ralph W

    2014-01-01

    The O. avara group of Oecetis is formally defined to include 4 described species, O. avara (Banks), O. disjuncta (Banks), O. elata Denning & Sykora, and O. metlacenis Bueno-Soria, and 15 new species. Oecetis marquesi Bueno-Soria, previously considered a member of the O. avara group, is treated as incertae sedis to species group, but is also redescribed and treated in the current work. New species described here (with their respective distributions) include: O. acciptrina (Costa Rica, Panama, Ecuador), O. agosta (Mexico), O. angularis (Guatemala to Ecuador), O. apache (SW USA), O. campana (Ecuador), O. constricta (Mexico to Ecuador, Venezuela, and Trinidad), O. houghtoni (North America), O. maritza (Costa Rica), O. mexicana (Mexico to Ecuador), O. patula (Guatemala, Nicaragua), O. protrusa (Mexico to Ecuador), O. sordida (Mexico, USA, Canada), O. tumida (Costa Rica), O. uncata (Costa Rica), and O. verrucula (Mexico to Costa Rica). A key to the species is also provided. PMID:24574849

  13. [The Medical Movement in Mexico 1964-1965: What's happening half a century later?].

    PubMed

    Gutiérrez-Samperio, César

    2016-01-01

    We reviewed the literature covering the medical movement in 1964-1965, which began on November 26, 1964 with the ISSSTE November 20 Hospital scholarship protest and the emergence of the Mexican Resident Interns Physicians Association (Asociación Mexicana de Médicos Residentes e Internos, AMMRI) and the Mexican Medical Alliance (Alianza de Médicos Mexicanos, AMM). We describe four work stoppages, two protest marches, the takeover of hospitals by the police, four interviews with Licenciado Gustavo Diaz Ordaz and his first presidential report. After that, attacks in the press, harassment, and repression provoked the weakening of the movement that ended in the AMM assembly on January 18, 1966. PMID:26927654

  14. Gynogenic plant regeneration from unpollinated flower explants of Eragrostis tef (Zuccagni) Trotter.

    PubMed

    Gugsa, Likyelesh; Sarial, Ashok K; Lörz, Horst; Kumlehn, Jochen

    2006-12-01

    Tef [Eragrostis tef (Zucc.) Trotter] is the most important cereal in Ethiopia. In its wild relative E. mexicana, regeneration of six green plants resulted from culture of 121 non-pollinated immature pistils. In the allotetraploid crop species tef, however, only callus and root formation was obtained by this method. By contrast, immature spikelets and panicle segments of E. tef proved amenable to gynogenic plant regeneration. Upon step-wise optimization of the protocol, efficient plant formation was achieved in all three cultivars tested. In cv. DZ-01-196, culture of 1305 immature spikelets resulted in formation of 159 green plants. Flow cytometric analysis revealed (di)haploid, triploid, tetraploid and octoploid regenerants, from which the vast majority was tetraploid. Tef-breeding programs will likely benefit substantially from efficient generation of true-breeding plants. PMID:16832620

  15. Extreme habitats as refuge from parasite infections? Evidence from an extremophile fish

    NASA Astrophysics Data System (ADS)

    Tobler, Michael; Schlupp, Ingo; García de León, Francisco J.; Glaubrecht, Matthias; Plath, Martin

    2007-05-01

    Living in extreme habitats typically requires costly adaptations of any organism tolerating these conditions, but very little is known about potential benefits that trade off these costs. We suggest that extreme habitats may function as refuge from parasite infections, since parasites can become locally extinct either directly, through selection by an extreme environmental parameter on free-living parasite stages, or indirectly, through selection on other host species involved in its life cycle. We tested this hypothesis in a small freshwater fish, the Atlantic molly ( Poecilia mexicana) that inhabits normal freshwaters as well as extreme habitats containing high concentrations of toxic hydrogen sulfide. Populations from such extreme habitats are significantly less parasitized by the trematode Uvulifer sp. than a population from a non-sulfidic habitat. We suggest that reduced parasite prevalence may be a benefit of living in sulfidic habitats.

  16. Genetic Variation of North American Triatomines (Insecta: Hemiptera: Reduviidae): Initial Divergence between Species and Populations of Chagas Disease Vector

    PubMed Central

    Espinoza, Bertha; Martínez-Ibarra, Jose Alejandro; Villalobos, Guiehdani; De La Torre, Patricia; Laclette, Juan Pedro; Martínez-Hernández, Fernando

    2013-01-01

    The triatomines vectors of Trypanosoma cruzi are principal factors in acquiring Chagas disease. For this reason, increased knowledge of domestic transmission of T. cruzi and control of its insect vectors is necessary. To contribute to genetic knowledge of North America Triatominae species, we studied genetic variations and conducted phylogenetic analysis of different triatomines species of epidemiologic importance. Our analysis showed high genetic variations between different geographic populations of Triatoma mexicana, Meccus longipennis, M. mazzottii, M. picturatus, and T. dimidiata species, suggested initial divergence, hybridation, or classifications problems. In contrast, T. gerstaeckeri, T. bolivari, and M. pallidipennis populations showed few genetics variations. Analysis using cytochrome B and internal transcribed spacer 2 gene sequences indicated that T. bolivari is closely related to the Rubrofasciata complex and not to T. dimidiata. Triatoma brailovskyi and T. gerstaeckeri showed a close relationship with Dimidiata and Phyllosoma complexes. PMID:23249692

  17. Molecular variability and evolution of the pectate lyase (pel-2) parasitism gene in cyst nematodes parasitizing different solanaceous plants.

    PubMed

    Geric Stare, Barbara; Fouville, Didier; Širca, Saša; Gallot, Aurore; Urek, Gregor; Grenier, Eric

    2011-02-01

    While pectate lyases are major parasitism factors in plant-parasitic nematodes, there is little information on the variability of these genes within species and their utility as pathotype or host range molecular markers. We have analysed polymorphisms of pectate lyase 2 (pel-2) gene, which degrades the unesterified polygalacturonate (pectate) of the host cell-wall, in the genus Globodera. Molecular variability of the pel-2 gene and the predicted protein was evaluated in populations of G. rostochiensis, G. pallida, G. "mexicana" and G. tabacum. Seventy eight pel-2 sequences were obtained and aligned. Point mutations were observed at 373 positions, 57% of these affect the coding part of the gene and produce 129 aa replacements. The observed polymorphism does not correlate either to the pathotypes proposed in potato cyst nematodes (PCN) or the subspecies described in tobacco cyst nematodes. The trees reveal a topology different from the admitted species topology as G. rostochiensis and G. pallida sequences are more similar to each other than to G. tabacum. Species-specific sites, potentially applicable for identification, and sites distinguishing PCN from tobacco cyst nematodes, were identified. As both G. rostochiensis and G. pallida display the same host range, but distinct from G. tabacum, which cannot parasitize potato plants, it is tempting to speculate that pel-2 genes polymorphism may be implicated in this adaptation, a view supported by the fact that no active pectate lyase 2 was found in G. "mexicana", a close relative of G. pallida that is unable to develop on cultivated potato varieties. PMID:21153407

  18. Molecular Diagnosis of Pathogenic Sporothrix Species

    PubMed Central

    Rodrigues, Anderson Messias; de Hoog, G. Sybren; de Camargo, Zoilo Pires

    2015-01-01

    Background Sporotrichosis is a chronic (sub)cutaneous infection caused by thermodimorphic fungi in the order, Ophiostomatales. These fungi are characterized by major differences in routes of transmission, host predilections, species virulence, and susceptibilities to antifungals. Sporothrix species emerge in the form of outbreaks. Large zoonoses and sapronoses are ongoing in Brazil and China, respectively. Current diagnostic methods based on morphology and physiology are inaccurate due to closely related phenotypes with overlapping components between pathogenic and non-pathogenic Sporothrix. There is a critical need for new diagnostic tools that are specific, sensitive, and cost-effective. Methodology We developed a panel of novel markers, based on calmodulin (CAL) gene sequences, for the large-scale diagnosis and epidemiology of clinically relevant members of the Sporothrix genus, and its relative, Ophiostoma. We identified specific PCR-based markers for S. brasiliensis, S. schenckii, S. globosa, S. mexicana, S. pallida, and O. stenoceras. We employed a murine model of disseminated sporotrichosis to optimize a PCR assay for detecting Sporothrix in clinical specimens. Results Primer-BLAST searches revealed candidate sequences that were conserved within a single species. Species-specific primers showed no significant homology with human, mouse, or microorganisms outside the Sporothrix genus. The detection limit was 10–100 fg of DNA in a single round of PCR for identifying S. brasiliensis, S. schenckii, S. globosa, S. mexicana, and S. pallida. A simple, direct PCR assay, with conidia as a source of DNA, was effective for rapid, low-cost genotyping. Samples from a murine model of disseminated sporotrichosis confirmed the feasibility of detecting S. brasiliensis and S. schenckii DNA in spleen, liver, lungs, heart, brain, kidney, tail, and feces of infected animals. Conclusions This PCR-based method could successfully detect and identify a single species in samples

  19. [Distribution and density of two holothurian species in Cubagua Island, Venezuela].

    PubMed

    Tagliafico, Alejandro; Rangel, María Salomé; Rago, Néstor

    2011-06-01

    Holothurian populations in the Caribbean have been significantly depleted in some areas, and others are still in danger to satisfy international markets. In Venezuela, this marine resource has been illegally exploited since the early 1990s. To contribute with their management and protection, this work analyzed the distribution, abundance and population structure of two commercial holothurians Isostichopus badionotus and Holothuria mexicana in Cubagua Island shores. A total of 52 stations were studied around the island during the year 2008. Surveys included visits to each station, for which a band transect of 50 m2 with four replicates were carried out. The results show that I. badionotus has a higher distribution on the East coast of the island, and is found over Thalassia testudinum beds or bivalve molluscs aggregations, with an average density of 0.011 ind/m2, C.I. 95% = 0.005-0.017, n = 122, and a general abundance estimated in 117 ind/ha. H. mexicana has been practically depleted (0.001 ind/m2 C.I. 95% = 0.0002-0.0013, 7.7 ind/ha, n = 8). We recommend the maintenance of the total ban adopted in 1997 to assure the protection of these species, mainly because no previous data supported the start of the fishery, and also, because the current densities are still very low when compared to other areas in Venezuela and the Caribbean. These are animals easy to catch, susceptible to overfishing, and with a very low recovery rate; thus, more control is required from the local authorities. PMID:21721242

  20. Delineation of Diverse Macrophage Activation Programs in Response to Intracellular Parasites and Cytokines

    PubMed Central

    Zhang, Shuyi; Kim, Charles C.; Batra, Sajeev; McKerrow, James H.; Loke, P'ng

    2010-01-01

    Background The ability to reside and proliferate in macrophages is characteristic of several infectious agents that are of major importance to public health, including the intracellular parasites Trypanosoma cruzi (the etiological agent of Chagas disease) and Leishmania species (etiological agents of Kala-Azar and cutaneous leishmaniasis). Although recent studies have elucidated some of the ways macrophages respond to these pathogens, the relationships between activation programs elicited by these pathogens and the macrophage activation programs elicited by bacterial pathogens and cytokines have not been delineated. Methodology/Principal Findings To provide a global perspective on the relationships between macrophage activation programs and to understand how certain pathogens circumvent them, we used transcriptional profiling by genome-wide microarray analysis to compare the responses of mouse macrophages following exposure to the intracellular parasites T. cruzi and Leishmania mexicana, the bacterial product lipopolysaccharide (LPS), and the cytokines IFNG, TNF, IFNB, IL-4, IL-10, and IL-17. We found that LPS induced a classical activation state that resembled macrophage stimulation by the Th1 cytokines IFNG and TNF. However, infection by the protozoan pathogen L. mexicana produced so few transcriptional changes that the infected macrophages were almost indistinguishable from uninfected cells. T. cruzi activated macrophages produced a transcriptional signature characterized by the induction of interferon-stimulated genes by 24 h post-infection. Despite this delayed IFN response by T. cruzi, the transcriptional response of macrophages infected by the kinetoplastid pathogens more closely resembled the transcriptional response of macrophages stimulated by the cytokines IL-4, IL-10, and IL-17 than macrophages stimulated by Th1 cytokines. Conclusions/Significance This study provides global gene expression data for a diverse set of biologically significant pathogens and

  1. Quantifying the severity of hurricanes on extinction probabilities of a primate population: Insights into "Island" extirpations.

    PubMed

    Ameca y Juárez, Eric I; Ellis, Edward A; Rodríguez-Luna, Ernesto

    2015-07-01

    Long-term studies quantifying impacts of hurricane activity on growth and trajectory of primate populations are rare. Using a 14-year monitored population of Alouatta palliata mexicana as a study system, we developed a modeling framework to assess the relative contribution of hurricane disturbance and two types of human impacts, habitat loss, and hunting, on quasi-extinction risk. We found that the scenario with the highest level of disturbance generated a 21% increase in quasi-extinction risk by 40 years compared to scenarios of intermediate disturbance, and around 67% increase relative to that found in low disturbance scenarios. We also found that the probability of reaching quasi-extinction due to human disturbance alone was below 1% by 40 years, although such scenarios reduced population size by 70%, whereas the risk of quasi-extinction ranged between 3% and 65% for different scenarios of hurricane severity alone, in absence of human impacts. Our analysis moreover found that the quasi-extinction risk driven by hunting and hurricane disturbance was significantly lower than the quasi-extinction risk posed by human-driven habitat loss and hurricane disturbance. These models suggest that hurricane disturbance has the potential to exceed the risk posed by human impacts, and, in particular, to substantially increase the speed of the extinction vortex driven by habitat loss relative to that driven by hunting. Early mitigation of habitat loss constituted the best method for reducing quasi-extinction risk: the earlier habitat loss is halted, the less vulnerable the population becomes to hurricane disturbance. By using a well-studied population of A. p. mexicana, we help understand the demographic impacts that extreme environmental disturbance can trigger on isolated populations of taxa already endangered in other systems where long-term demographic data are not available. For those experiencing heavy anthropogenic pressure and lacking sufficiently evolved coping

  2. Antimicrobial and antioxidant activities of plants from northeast of Mexico.

    PubMed

    Salazar-Aranda, Ricardo; Pérez-López, Luis Alejandro; López-Arroyo, Joel; Alanís-Garza, Blanca Alicia; Waksman de Torres, Noemí

    2011-01-01

    Traditional medicine has a key role in health care worldwide. Obtaining scientific information about the efficacy and safety of the plants from our region is one of the goals of our research group. In this report, 17 plants were selected and collected in different localities from northeast Mexico. The dried plants were separated into leaves, flowers, fruit, stems, roots and bark. Each part was extracted with methanol, and 39 crude extracts were prepared. The extracts were tested for their antimicrobial activity using three Gram-negative bacterial strains (Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii), three Gram-positive bacterial strains (Enterococcus faecalis and two Staphylococcus aureus strains), and seven clinically isolated yeasts (Candida albicans, C. krusei, C. tropicalis, C. parapsilosis and C. glabrata); their antioxidant activity was tested using a DPPH free radical assay. No activity against Gram-negative bacteria was observed with any extract up to the maximum concentration tested, 1000 μg ml(-1). We report here for the first time activity of Ceanothus coeruleus against S. aureus (flowers, minimal inhibitory concentration (MIC) 125 μg ml(-1)), C. glabrata (MICs 31.25 μg ml(-1)) and C. parapsilosis (MICs between 31.25 and 125 μg ml(-1)); Chrysanctinia mexicana against C. glabrata (MICs 31.25 μg ml(-1)); Colubrina greggii against E. faecalis (MICs 250 μg ml(-1)) and Cordia boissieri against C. glabrata (MIC 125 μg ml(-1)). Furthermore, this is the first report about antioxidant activity of extracts from Ceanothus coeruleus, Chrysanctinia mexicana, Colubrina greggii and Cyperus alternifolius. Some correlation could exist between antioxidant activity and antiyeast activity against yeasts in the species Ceanothus coeruleus, Schinus molle, Colubrina greggii and Cordia boissieri. PMID:19770266

  3. Antimicrobial and Antioxidant Activities of Plants from Northeast of Mexico

    PubMed Central

    Salazar-Aranda, Ricardo; Pérez-López, Luis Alejandro; López-Arroyo, Joel; Alanís-Garza, Blanca Alicia; Waksman de Torres, Noemí

    2011-01-01

    Traditional medicine has a key role in health care worldwide. Obtaining scientific information about the efficacy and safety of the plants from our region is one of the goals of our research group. In this report, 17 plants were selected and collected in different localities from northeast Mexico. The dried plants were separated into leaves, flowers, fruit, stems, roots and bark. Each part was extracted with methanol, and 39 crude extracts were prepared. The extracts were tested for their antimicrobial activity using three Gram-negative bacterial strains (Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii), three Gram-positive bacterial strains (Enterococcus faecalis and two Staphylococcus aureus strains), and seven clinically isolated yeasts (Candida albicans, C. krusei, C. tropicalis, C. parapsilosis and C. glabrata); their antioxidant activity was tested using a DPPH free radical assay. No activity against Gram-negative bacteria was observed with any extract up to the maximum concentration tested, 1000 μg ml−1. We report here for the first time activity of Ceanothus coeruleus against S. aureus (flowers, minimal inhibitory concentration (MIC) 125 μg ml−1), C. glabrata (MICs 31.25 μg ml−1) and C. parapsilosis (MICs between 31.25 and 125 μg ml−1); Chrysanctinia mexicana against C. glabrata (MICs 31.25 μg ml−1); Colubrina greggii against E. faecalis (MICs 250 μg ml−1) and Cordia boissieri against C. glabrata (MIC 125 μg ml−1). Furthermore, this is the first report about antioxidant activity of extracts from Ceanothus coeruleus, Chrysanctinia mexicana, Colubrina greggii and Cyperus alternifolius. Some correlation could exist between antioxidant activity and antiyeast activity against yeasts in the species Ceanothus coeruleus, Schinus molle, Colubrina greggii and Cordia boissieri. PMID:19770266

  4. Effect of Ambient Temperature on the Clinical Manifestations of Experimental Diffuse Cutaneous Leishmaniasis in a Rodent Model

    PubMed Central

    Quiñonez-Díaz, Laura; Mancilla-Ramírez, Javier; Avila-García, Miroslava; Ortiz-Avalos, Juana; Berron, Angélica; González, Susana; Paredes, Yuriria

    2012-01-01

    Abstract Dermal species of Leishmania have a relatively broad temperature range for optimal growth in vitro, with temperature differences accompanied by a form change. This suggests that when the host is living in moderate temperatures (22°C), infection may proceed at temperatures lower than those that occur in tropical regions (32°C), and a different clinical expression of the disease due to a different parasitic form may result. The aim of this study was to investigate the effect of environmental temperature on the clinical expression of the disease. BALB/C mice infected with Leishmania mexicana were housed at 32°±2°C or 22°±1°C, and assessed for the development of inflammation and the presence of parasites in organs using PCR and immunohistology. The clinical expression of leishmaniasis at 32°C included inflammation at the site of inoculation with swelling of the nose and tail, whereas at 22°C, up to 50% of the infected mice developed dry exfoliative dermatitis with alopecia on the dorsum. In both cases, parasite colonization was confirmed in the skin, with parasites at more external locations at 22°C. Parasite visceralization was confirmed in all internal organs and glands in both cases based on PCR and immunohistology. In conclusion, the clinical expression of diffuse leishmaniasis by Leishmania mexicana in laboratory mice is modified by temperature, from nodular inflammation at 32°C, to dry exfoliative dermatitis and alopecia at 22°C, with parasite visceralization in both cases. PMID:22651383

  5. Life Course Experiences, Pain and Suffering: A Case Study of an Older Mexican American Woman with Mobility Impairment

    PubMed Central

    Walker, Janiece L.; Harrison, Tracie C.; Hendrickson, Sherry G.

    2012-01-01

    There is a dearth of literature examining how adversity shapes the experiences of pain and/or suffering in a middle aged Mexican American women. The purpose of this qualitative descriptive study was to understand pain and suffering from a life course perspective as described by a Mexican American woman aging with early onset mobility impairment. This Hispanic woman experienced episodes of abuse and rejection over the life course, which may have significantly influenced her pain and suffering experience in adulthood. This adds to the literature on how adversity influences later life pain experience and provides insight on why pharmacological treatment alone may not be as successful as a holistic intervention. Hay escasez de literatura que examine cómo la adversidad da forma a las experiencias de dolor y / o sufrimiento en mujeres mexicana-americana mayores de edad. El propósito de este estudio descriptivo cualitativo fue comprender el dolor y el sufrimiento desde una perspectiva de ciclo de vida como descrito por una mujer mexicana-americana envejeciendo con inicio temprano de deterioro de movilidad. Esta Latina experimentó episodios de maltrato y rechazo, que se percibió haber afectado su experiencia de dolor y sufrimiento luego como adulta. Este estudio aumenta la literatura sobre cómo la adversidad influya la experiencia de dolor más tarde en la vida, y nos permite comprender mejor como el tratamiento farmacológico por sí solo no es tan exitoso como pueda ser una intervención integral. PMID:24830728

  6. The effect of chemical processing on the δ 13C value of plant tissue

    NASA Astrophysics Data System (ADS)

    Van de Water, Peter K.

    2002-04-01

    The effect of standard processing techniques on the δ 13C value of plant tissue was tested using species representing the three photosynthetic pathways, including angiosperms and gymnosperms within the C 3 taxonomic division. The species include Cowania mexicana (C 3 angiosperm), Juniperus osteosperma (C 3 gymnosperm), Opuntia spp. (crassulacean acid metabolism [CAM] angiosperm), and Atriplex canescens (C 4 angiosperm). Each species is represented by 5 plants collected at two different sites, for a total of 10 samples. The samples were processed to whole plant tissue, holocellulose, α-cellulose, and nitrocellulose. An additional process was added with the discovery of residual Ca-oxalate crystals in holocellulose samples. Both C 3 species show δ 13C values becoming 13C enriched with increased processing. The CAM representative shows the opposite trend, with 13C depletion during the progression of treatments. The greatest range of values and most inconsistent trends occur in the C 4 representative. Removal of the Ca-oxalate fraction resulted in different mean weight percentages and δ 13C values among the species. Calculated δ 13C values of the Ca-oxalate crystals show depletion from the tissue values in the two C 3 species and enrichment in the C 4 and CAM representatives. The C. mexicana samples show the greatest change between the tissue and Ca-oxalates (7.3‰) but the least mean weight percentage (11%), whereas A. canescens shows the greatest overall change, with a -2.8‰ isotopic shift and over 48% mean weight percentage. Variability within the samples undergoing each treatment remained relatively unchanged even with increased cellulose purity. This paper provides estimates of isotopic offsets necessary to correct from one treatment to another. Significant differences in δ 13C among different treatments confirm the need to state the tissue fraction analyzed when reporting δ 13C results.

  7. Phylogeny and biogeography of the Poecilia sphenops species complex (Actinopterygii, Poeciliidae) in Central America.

    PubMed

    Alda, Fernando; Reina, Ruth G; Doadrio, Ignacio; Bermingham, Eldredge

    2013-03-01

    We inferred the phylogenetic relationships among members of the Poecilia sphenops species complex to resolve the colonization process and radiation of this group in Central America. We analyzed 2550 base pairs (bp) of mitochondrial DNA (mtDNA), including ATP synthase 6 and 8, cytochrome oxidase subunit I and NADH dehydrogenase subunit 2 genes, and 906bp of the nuclear S7 ribosomal protein of 86 ingroup individuals from 61 localities spanning most of its distribution from Mexico to Panama. Our mitochondrial data rendered a well-supported phylogeny for the P. sphenops complex that differed with the nuclear data set topology, which did not recover the monophyly of the P. mexicana mitochondrial lineage. Coalescent-based simulations tests indicated that, although hybridization cannot be completely ruled out, this incongruence is most likely due to incomplete lineage sorting in this group, which also showed the widest geographic distribution. A single colonization event of Central America from South America was estimated to have occurred between the early Paleocene and Oligocene (53-22millionyears ago). Subsequently, two largely differentiated evolutionary lineages diverged around the Early Oligocene-Miocene (38-13million years ago), which are considered two separate species complexes: P. sphenops and P. mexicana, which can also be distinguished by their tricuspid and unicuspid inner jaw teeth, respectively. Ultimately, within lineage diversification occurred mainly during the Miocene (22-5million years ago). All major cladogenetic events predated the final closure of the Isthmus of Panama. The allopatric distribution of lineages together with the long basal internodes suggest that vicariance and long term isolations could be the main evolutionary forces promoting radiation in this group, although dispersal through water barriers might also have occurred. Lastly, our results suggest the need to review the current species distribution and taxonomy of the P. sphenops

  8. Predator-induced changes of female mating preferences: innate and experiential effects

    PubMed Central

    2011-01-01

    Background In many species males face a higher predation risk than females because males display elaborate traits that evolved under sexual selection, which may attract not only females but also predators. Females are, therefore, predicted to avoid such conspicuous males under predation risk. The present study was designed to investigate predator-induced changes of female mating preferences in Atlantic mollies (Poecilia mexicana). Males of this species show a pronounced polymorphism in body size and coloration, and females prefer large, colorful males in the absence of predators. Results In dichotomous choice tests predator-naïve (lab-reared) females altered their initial preference for larger males in the presence of the cichlid Cichlasoma salvini, a natural predator of P. mexicana, and preferred small males instead. This effect was considerably weaker when females were confronted visually with the non-piscivorous cichlid Vieja bifasciata or the introduced non-piscivorous Nile tilapia (Oreochromis niloticus). In contrast, predator experienced (wild-caught) females did not respond to the same extent to the presence of a predator, most likely due to a learned ability to evaluate their predators' motivation to prey. Conclusions Our study highlights that (a) predatory fish can have a profound influence on the expression of mating preferences of their prey (thus potentially affecting the strength of sexual selection), and females may alter their mate choice behavior strategically to reduce their own exposure to predators. (b) Prey species can evolve visual predator recognition mechanisms and alter their mate choice only when a natural predator is present. (c) Finally, experiential effects can play an important role, and prey species may learn to evaluate the motivational state of their predators. PMID:21726456

  9. Prickly Poppies Can Get Pricklier: Ontogenetic Patterns in the Induction of Physical Defense Traits

    PubMed Central

    Hoan, Ryan P.; Ormond, Rhys A.; Barton, Kasey E.

    2014-01-01

    Plant ontogeny is a common source of variation in defense and herbivory. Yet, few studies have investigated how the induction of physical defense traits changes across plant ontogeny. Physical defense traits are costly to produce, and thus, it was predicted that induction as a cost-saving strategy would be particularly favorable for seedlings, leading to ontogenetic declines in the inducibility of these traits. We tested for induction of three different physical defense traits (prickles, latex and leaf toughness) in response to mechanical defoliation and jasmonic acid application using prickly poppies (Argemone glauca and A. mexicana, Papaveraceae) as a model system. Genetic variation in the induction of physical defenses was tested using maternal sib-ships sampled from multiple populations. Both species induced higher densities of laminar prickles, although the magnitude of induction was much higher in the endemic Hawaiian prickly poppy, A. glauca, than in the cosmopolitan A. mexicana. The magnitude of prickle induction was also higher in young compared to older juvenile plant stages in A. glauca, demonstrating a strong role of ontogeny. Neither latex exudation nor leaf toughness was induced in either species. Although significant genetic variation was detected within and among populations for constitutive expression of physical defense traits in Argemone, there was no evidence for genetic variation in the induction of these traits. This study provides the first evidence for the induction of physical defenses in prickly poppies, emphasizing how an ontogenetically explicit framework can reveal new insights into plant defense. Moreover, this study illustrates how sister species comparisons between island vs. continental plants can provide new insights into plant functional and evolutionary ecology, highlighting a fruitful area for future research on more species pairs. PMID:24802133

  10. The Rediscovery of a Long Described Species Reveals Additional Complexity in Speciation Patterns of Poeciliid Fishes in Sulfide Springs

    PubMed Central

    Palacios, Maura; Arias-Rodriguez, Lenin; Plath, Martin; Eifert, Constanze; Lerp, Hannes; Lamboj, Anton; Voelker, Gary; Tobler, Michael

    2013-01-01

    The process of ecological speciation drives the evolution of locally adapted and reproductively isolated populations in response to divergent natural selection. In Southern Mexico, several lineages of the freshwater fish species of the genus Poecilia have independently colonized toxic, hydrogen sulfide-rich springs. Even though ecological speciation processes are increasingly well understood in this system, aligning the taxonomy of these fish with evolutionary processes has lagged behind. While some sulfide spring populations are classified as ecotypes of Poecilia mexicana, others, like P. sulphuraria, have been described as highly endemic species. Our study particularly focused on elucidating the taxonomy of the long described sulfide spring endemic, Poecilia thermalis Steindachner 1863, and investigates if similar evolutionary patterns of phenotypic trait divergence and reproductive isolation are present as observed in other sulfidic species of Poecilia. We applied a geometric morphometric approach to assess body shape similarity to other sulfidic and non-sulfidic fish of the genus Poecilia. We also conducted phylogenetic and population genetic analyses to establish the phylogenetic relationships of P. thermalis and used a population genetic approach to determine levels of gene flow among Poecilia from sulfidic and non-sulfidic sites. Our results indicate that P. thermalis' body shape has evolved in convergence with other sulfide spring populations in the genus. Phylogenetic analyses placed P. thermalis as most closely related to one population of P. sulphuraria, and population genetic analyses demonstrated that P. thermalis is genetically isolated from both P. mexicana ecotypes and P. sulphuraria. Based on these findings, we make taxonomic recommendations for P. thermalis. Overall, our study verifies the role of hydrogen sulfide as a main factor shaping convergent, phenotypic evolution and the emergence of reproductive isolation between Poecilia populations

  11. IFN-gamma expression is up-regulated by peripheral blood mononuclear cells (PBMC) from non-exposed dogs upon Leishmania chagasi promastigote stimulation in vitro.

    PubMed

    Rodrigues, Cleusa Alves Theodore; Batista, Luís Fábio da Silva; Filho, Roberto Santos Teixeira; Santos, Claire da Silva; Pinheiro, Cristiane Garboggini; Almeida, Taís Fontoura de; Freitas, Luiz Antônio Rodrigues de; Veras, Patrícia Sampaio Tavares

    2009-02-15

    While the response to Leishmania spp. is well characterized in mice and humans, much less is known concerning the canine immune response, particularly soon after exposure to the parasite. Early events are considered to be a determinant of infection outcome. To investigate the dog's early immune response to L. chagasi, an in vitro priming system (PIV) using dog naïve PBMC was established. Until now, dog PIV immune response to L. chagasi has not been assessed. We co-cultivated PBMC primarily stimulated with L. chagasi in vitro with autologous infected macrophages and found that IFN-gamma mRNA is up-regulated in these cells compared to control unstimulated cells. IL-4 and IL-10 mRNA expression by L. chagasi-stimulated PBMC was similar to control unstimulated PBMC when incubated with infected macrophages. Surprisingly, correlation studies showed that a lower IFN-gamma/IL-4 expression ratio correlated with a lower percentage of infection. We propose that the direct correlation between IFN-gamma/IL-4 ratio and parasite load is dependent on the higher correlation of both IFN-gamma and IL-4 expression with lower parasite infection. This PIV system was shown to be useful in evaluating the dog immune response to L. chagasi, and results indicate that a balance between IFN-gamma and IL-4 is associated with control of parasite infection in vitro. PMID:19054575

  12. Description of Globodera ellingtonae n. sp. (Nematoda: Heteroderidae) from Oregon.

    PubMed

    Handoo, Zafar A; Carta, Lynn K; Skantar, Andrea M; Chitwood, David J

    2012-03-01

    A new species of cyst nematode, Globodera ellingtonae, is described from soil collected from a field in Oregon. Second-stage juveniles (J2) of the species are characterized by body length of 365-515 μm, stylet length of 19-22.5 μm, basal knobs rounded posteriorly and pointed anteriorly, tail 39-55 μm, hyaline tail terminus 20-32.5 μm, and tail tapering uniformly but abruptly narrowing and constricted near the posterior third of the hyaline portion, ending with a peg-like, finely rounded to pointed terminus. Cysts are spherical to sub-spherical, dark to light brown and circumfenestrate and cyst wall pattern is ridge-like with heavy punctations. Males have a stylet length of 21-25 μm and spicule length of 30-37 μm with a pointed thorn-like tip. Females have a stylet length of 20-22.5 μm, one head annule and labial disc, heavy punctations on the cuticle, and short vulval slit 7.5-8 μm long. Morphologically this new, round-cyst species differs from the related species G. pallida, G. rostochiensis, G. tabacum complex and G. mexicana by its distinctive J2 tail, and by one or another of the following: shorter mean stylet length in J2, females and males; number of refractive bodies in the hyaline tail terminus of J2; cyst morphology including Granek's ratio; number of cuticular ridges between the anus and vulva; and in the shape and length of spicules in males. Its relationship to these closely related species are discussed. Based upon analysis of ribosomal internal transcribed spacer (ITS) sequences, G. ellingtonae n. sp. is distinct from G. pallida, G. rostochiensis, G. tabacum and G. mexicana. Bayesian and Maximum Parsimony analysis of cloned ITS rRNA gene sequences indicated three clades, with intraspecific variability as high as 2.8%. In silico analysis revealed ITS restriction fragment length polymorphisms for enzymes Bsh 1236I, Hinf I, and Rsa I that overlap patterns for other Globodera species. PMID:23483076

  13. Description of Globodera ellingtonae n. sp. (Nematoda: Heteroderidae) from Oregon

    PubMed Central

    Handoo, Zafar A.; Carta, Lynn K.; Skantar, Andrea M.; Chitwood, David J.

    2012-01-01

    A new species of cyst nematode, Globodera ellingtonae, is described from soil collected from a field in Oregon. Second-stage juveniles (J2) of the species are characterized by body length of 365-515 μm, stylet length of 19-22.5 μm, basal knobs rounded posteriorly and pointed anteriorly, tail 39-55 μm, hyaline tail terminus 20-32.5 μm, and tail tapering uniformly but abruptly narrowing and constricted near the posterior third of the hyaline portion, ending with a peg-like, finely rounded to pointed terminus. Cysts are spherical to sub-spherical, dark to light brown and circumfenestrate and cyst wall pattern is ridge-like with heavy punctations. Males have a stylet length of 21-25 μm and spicule length of 30-37 μm with a pointed thorn-like tip. Females have a stylet length of 20-22.5 μm, one head annule and labial disc, heavy punctations on the cuticle, and short vulval slit 7.5-8 μm long. Morphologically this new, round-cyst species differs from the related species G. pallida, G. rostochiensis, G. tabacum complex and G. mexicana by its distinctive J2 tail, and by one or another of the following: shorter mean stylet length in J2, females and males; number of refractive bodies in the hyaline tail terminus of J2; cyst morphology including Granek’s ratio; number of cuticular ridges between the anus and vulva; and in the shape and length of spicules in males. Its relationship to these closely related species are discussed. Based upon analysis of ribosomal internal transcribed spacer (ITS) sequences, G. ellingtonae n. sp. is distinct from G. pallida, G. rostochiensis, G. tabacum and G. mexicana. Bayesian and Maximum Parsimony analysis of cloned ITS rRNA gene sequences indicated three clades, with intraspecific variability as high as 2.8%. In silico analysis revealed ITS restriction fragment length polymorphisms for enzymes Bsh 1236I, Hinf I, and Rsa I that overlap patterns for other Globodera species. PMID:23483076

  14. Sequence Evolution and Expression of the Androgen Receptor and Other Pathway-Related Genes in a Unisexual Fish, the Amazon Molly, Poecilia formosa, and Its Bisexual Ancestors

    PubMed Central

    Zhu, Fangjun; Schlupp, Ingo; Tiedemann, Ralph

    2016-01-01

    The all-female Amazon molly (Poecilia formosa) originated from a single hybridization of two bisexual ancestors, Atlantic molly (Poecilia mexicana) and sailfin molly (Poecilia latipinna). As a gynogenetic species, the Amazon molly needs to copulate with a heterospecific male, but the genetic information of the sperm-donor does not contribute to the next generation, as the sperm only acts as the trigger for the diploid eggs’ embryogenesis. Here, we study the sequence evolution and gene expression of the duplicated genes coding for androgen receptors (ars) and other pathway-related genes, i.e., the estrogen receptors (ers) and cytochrome P450, family19, subfamily A, aromatase genes (cyp19as), in the Amazon molly, in comparison to its bisexual ancestors. Mollies possess–as most other teleost fish—two copies of the ar, er, and cyp19a genes, i.e., arα/arβ, erα/erβ1, and cyp19a1 (also referred as cyp19a1a)/cyp19a2 (also referred to as cyp19a1b), respectively. Non-synonymous single nucleotide polymorphisms (SNPs) among the ancestral bisexual species were generally predicted not to alter protein function. Some derived substitutions in the P. mexicana and one in P. formosa are predicted to impact protein function. We also describe the gene expression pattern of the ars and pathway-related genes in various tissues (i.e., brain, gill, and ovary) and provide SNP markers for allele specific expression research. As a general tendency, the levels of gene expression were lowest in gill and highest in ovarian tissues, while expression levels in the brain were intermediate in most cases. Expression levels in P. formosa were conserved where expression did not differ between the two bisexual ancestors. In those cases where gene expression levels significantly differed between the bisexual species, P. formosa expression was always comparable to the higher expression level among the two ancestors. Interestingly, erβ1 was expressed neither in brain nor in gill in the analyzed

  15. Systematics of Disakisperma (Poaceae, Chloridoideae, Chlorideae)

    PubMed Central

    Snow, Neil; Peterson, Paul M.; Romaschenko, Konstantin

    2013-01-01

    Abstract Disakisperma Steud. is a genus of four predominantly perennial C4 (NAD-ME) species in the Americas, Africa, and Asia. Its species previously were treated in Eleusine, Eragrostis, Coelachyrum, Cypholepis, Leptochloa, or Diplachne by nearly all authors.It includes the widespread North and South American amphitropical disjunct Disakisperma dubium (type of the genus), Disakisperma eleusine from southern Africa, Disakisperma obtusiflorum from central and northern Africa to southern Asia, and Disakisperma yemenicum, comb. nov. from eastern and southern Africa to Yemen. This paper provides a key to the species, geographic distributions, descriptions, including comments on the anatomy of leaves, stems, lemmatal micromorphology, a phylogram based on five molecular markers, and discussions of chromosome numbers. The species are rarely, if at all, known outside of their native ranges and are unlikely to become aggressively invasive. All species are considered Least Concern following IUCN guidelines. Lectotypes are designated for Diplachne dubia var. pringleana Kuntze, Disakisperma mexicana Steud., Eragrostis yemenica Schweinf., and Leptochloa appletonii Stapf. PMID:24194669

  16. Plants used in Guatemala for the treatment of gastrointestinal disorders. 1. Screening of 84 plants against enterobacteria.

    PubMed

    Caceres, A; Cano, O; Samayoa, B; Aguilar, L

    1990-08-01

    Gastrointestinal disorders are important causes of morbidity in developing countries. Natural healing is the traditional way of treating these diseases in Guatemala. Ethnobotanical surveys and literature reviews showed that 385 plants from 95 families are used in Guatemala for the treatment of gastrointestinal disorders. The activity of 84 of the most commonly used plants was screened in vitro against five enterobacteria pathogenic to man (enteropathogenic Escherichia coli, Salmonella enteritidis, Salmonella typhi, Shigella dysenteriae and Shigella flexneri). Results indicate that 34 (40.48%) plants inhibit one or more of the enterobacteria tested. The most commonly inhibited bacterium was S. typhi (33.73%) and the most resistant was E. coli (7.35%). The plants of American origin which exhibited the best antibacterial activity were: Byrsonima crassifolia, Diphysa robinioides, Gnaphalium stramineum, Guazuma ulmifolia, Psidium guajava, Sambucus mexicana, Simarouba glauca, Smilax lundelii, Spondias purpurea and Tagetes lucida. These results indicate a scientific basis for use of these medicinal plants for attacking enterobacterial infections in man. PMID:2214824

  17. Avian Conservation Practices Strengthen Ecosystem Services in California Vineyards

    PubMed Central

    Jedlicka, Julie A.; Greenberg, Russell; Letourneau, Deborah K.

    2011-01-01

    Insectivorous Western Bluebirds (Sialia mexicana) occupy vineyard nest boxes established by California winegrape growers who want to encourage avian conservation. Experimentally, the provision of available nest sites serves as an alternative to exclosure methods for isolating the potential ecosystem services provided by foraging birds. We compared the abundance and species richness of avian foragers and removal rates of sentinel prey in treatments with songbird nest boxes and controls without nest boxes. The average species richness of avian insectivores increased by over 50 percent compared to controls. Insectivorous bird density nearly quadrupled, primarily due to a tenfold increase in Western Bluebird abundance. In contrast, there was no significant difference in the abundance of omnivorous or granivorous bird species some of which opportunistically forage on grapes. In a sentinel prey experiment, 2.4 times more live beet armyworms (Spodoptera exigua) were removed in the nest box treatment than in the control. As an estimate of the maximum foraging services provided by insectivorous birds, we found that larval removal rates measured immediately below occupied boxes averaged 3.5 times greater than in the control. Consequently the presence of Western Bluebirds in vineyard nest boxes strengthened ecosystem services to winegrape growers, illustrating a benefit of agroecological conservation practices. Predator addition and sentinel prey experiments lack some disadvantages of predator exclusion experiments and were robust methodologies for detecting ecosystem services. PMID:22096555

  18. Partial nitrification using aerobic granules in continuous-flow reactor: rapid startup.

    PubMed

    Wan, Chunli; Sun, Supu; Lee, Duu-Jong; Liu, Xiang; Wang, Li; Yang, Xue; Pan, Xiangliang

    2013-08-01

    This study applied a novel strategy to rapid startup of partial nitrification in continuous-flow reactor using aerobic granules. Mature aerobic granules were first cultivated in a sequencing batch reactor at high chemical oxygen demand in 16 days. The strains including the Pseudoxanthomonas mexicana strain were enriched in cultivated granules to enhance their structural stability. Then the cultivated granules were incubated in a continuous-flow reactor with influent chemical oxygen deamnad being stepped decreased from 1,500 ± 100 (0-19 days) to 750 ± 50 (20-30 days), and then to 350 ± 50 mg l(-1) (31-50 days); while in the final stage 350 mg l(-1) bicarbonate was also supplied. Using this strategy the ammonia-oxidizing bacterium, Nitrosomonas europaea, was enriched in the incubated granules to achieve partial nitrification efficiency of 85-90% since 36 days and onwards. The partial nitrification granules were successfully harvested after 52 days, a period much shorter than those reported in literature. PMID:23751489

  19. In Silico Elucidation and Inhibition Studies of Selected Phytoligands Against Mitogen-Activated Protein Kinases of Protozoan Parasites.

    PubMed

    Gupta, Chhedi Lal; Akhtar, Salman; Kumar, Nilesh; Ali, Jasarat; Pathak, Neelam; Bajpai, Preeti

    2016-03-01

    Parasitic MAPKs exhibiting significant divergence with humans and playing an imperative role in parasitic metabolic activities have been exploited from several years as important targets for development of novel therapeutics. In addition, the emergence of the drug-resistant variants of parasitic diseases in the recent years has aroused a great need for the development of potent inhibitors against them. In the present study, we selected the metabolically active MAPKs LmxMPK4, PfMAP2 and TbMAPK5 of the three parasitic protozoans Leishmania mexicana, Plasmodium falciparum and Trypanosoma brucei, respectively. The homology modeling technique was used to develop the 3D structures of these proteins, and the same was validated by PROCHECK, ERRAT, ProQ and ProSA web servers to check the reliability. Ten phytoligands were employed for molecular docking studies with these proteins to search for potent phytoligand as a broad spectrum inhibitor. In this regard, two phytoligands (aspidocarpine for LmxMPK4 and TbMAPK5 and cubebin for PfMAP2) were found to be more effective inhibitors, in terms of robust binding energy, strong inhibition constant and better interactions between protein-ligand complexes. Furthermore, predicted ADME and toxicity properties suggested that these identified phytoligands exhibited comparable results to control drugs potentiating them as persuasive therapeutic agents for Leishmania, Trypanosoma and Plasmodium sp. PMID:26264054

  20. Infectious disease survey of gemsbok in New Mexico

    USGS Publications Warehouse

    Bender, L.C.; Li, H.; Thompson, B.C.; Morrow, P.C.; Valdez, R.

    2003-01-01

    Exotic wildlife can introduce new diseases or act as reservoirs of endemic diseases. On White Sands Missile Range, New Mexico (USA), significant declines in populations of native ungulates generally correspond to increases in range and population density of the exotic gemsbok (Oryx gazella gazella), introduced beginning in 1969. We surveyed gemsbok in 2001 for exposure to a variety of diseases potentially important for native ungulates. High seroprevalence was found for malignant catarrhal fever virus (49 [98%] of 50 sera; 43 [96%] of 45 plasma samples), bluetongue virus (48 [96%] of 50), bovine respiratory syncytial virus (33 [66%] of 50), and parainfluenza-3 virus (10 [20%] of 50). Low numbers of Nematodirus spp. eggs in a few individuals were the only parasites detected in gemsbok. Exposure to the above diseases in gemsbok is of interest to managers because of potential implications for recovery of desert bighorn sheep (Ovis canadensis mexicana) and desert mule deer (Odocoileus hemionus crooki) in the White Sands area because each has been implicated in mortality in these species either in the White Sands area or elsewhere in the western/southwestern United States. ?? Wildlife Disease Association 2003.

  1. Relation of desert pupfish abundance to selected environmental variables in natural and manmade habitats in the Salton Sea basin

    USGS Publications Warehouse

    Martin, B.A.; Saiki, M.K.

    2005-01-01

    We assessed the relation between abundance of desert pupfish, Cyprinodon macularius, and selected biological and physicochemical variables in natural and manmade habitats within the Salton Sea Basin. Field sampling in a natural tributary, Salt Creek, and three agricultural drains captured eight species including pupfish (1.1% of the total catch), the only native species encountered. According to Bray-Curtis resemblance functions, fish species assemblages differed mostly between Salt Creek and the drains (i.e., the three drains had relatively similar species assemblages). Pupfish numbers and environmental variables varied among sites and sample periods. Canonical correlation showed that pupfish abundance was positively correlated with abundance of western mosquitofish, Gambusia affinis, and negatively correlated with abundance of porthole livebearers, Poeciliopsis gracilis, tilapias (Sarotherodon mossambica and Tilapia zillii), longjaw mudsuckers, Gillichthys mirabilis, and mollies (Poecilia latipinnaandPoecilia mexicana). In addition, pupfish abundance was positively correlated with cover, pH, and salinity, and negatively correlated with sediment factor (a measure of sediment grain size) and dissolved oxygen. Pupfish abundance was generally highest in habitats where water quality extremes (especially high pH and salinity, and low dissolved oxygen) seemingly limited the occurrence of nonnative fishes. This study also documented evidence of predation by mudsuckers on pupfish. These findings support the contention of many resource managers that pupfish populations are adversely influenced by ecological interactions with nonnative fishes. ?? Springer 2005.

  2. Antigenic Diversity and Distribution of Rabies Virus in Mexico

    PubMed Central

    Velasco-Villa, Andrés; Gómez-Sierra, Mauricio; Hernández-Rodríguez, Gustavo; Juárez-Islas, Victor; Meléndez-Félix, Alejandra; Vargas-Pino, Fernando; Velázquez-Monroy, Oscar; Flisser, Ana

    2002-01-01

    Rabies remains a public health problem in the Americas because of the great diversity of wild reservoirs that maintain the virus in nature. Here we report the antigenic characterization of 254 rabies viruses isolated from 148 nonreservoir and 106 reservoir hosts collected in 27 states of Mexico. Nine out of 11 antigenic variants previously reported in the United States were detected in Mexico by using the limited panel of monoclonal antibodies donated by the Centers for Disease Control and Prevention. Some rabies virus variants were isolated from their natural reservoirs, which were also taxonomically identified. Terrestrial reservoirs included stray dogs with V1, Urocyon cineroargenteus (gray foxes) with V7, and two subspecies of Spilogale putorius (spotted skunks) with different viral variants (V8 and V10). Aerial hosts included Tadarida brasiliensis mexicana and Desmodus rotundus, which harbored V9 and V4 and harbored V11, respectively. All variants, with the exception of V9, were isolated from nonreservoir hosts, while V3, V4, and V5 were not isolated from their natural reservoirs but only from livestock. Rabies virus antigenic typing allowed us to determine rabies reservoirs and their distribution in Mexico, data which will probably improve prevention and control of the illness in humans and in the reservoir hosts. PMID:11880422

  3. Common environmental allergens causing respiratory allergy in India.

    PubMed

    Singh, A B; Kumar, Pawan

    2002-03-01

    Respiratory allergy affects all age groups but the children are the worst affected by the respiratory allergy. Bioparticles from different biological sources are the main cause of allergy. Pollen grains, fungal spores, insect and other materials of biological origin form the most important allergen load in the air. For the efficient diagnosis of the allergy and its effective treatment it is very important to know about the prevalence, seasonal and annual variations of aeroallergens of the area. India being the climatically diversed country, there is diversity in the flora and fauna of different parts of the country. Atmospheric surveys carried out in different parts of India reveal that, Alanus nitida, Amarantus spinosus, Argemone mexicana Cocos nucifera, Betula utilis, Borasus flabellifer, Caraica papaya, Cedrus deodara, Cassia fistula, Parthenium, Chenopodium album, Dodonaea viscosa, Malotus phillipensis, Plantago ovata, Prosopis juliflora, Ricinus communis, Holoptelea intergifolia are the allergenically important pollens of the country. Among the fungal aeroallergens, Alternaria, Candida aibieans, Aspergillus versicolor, Aspergillus terreus, Aspergillus japonicus, Cladosporium cladosporoides, Fusarium roseum, Ganoderma lucidum,Neurospora sitophila Helminthosporium, Ustilago trtici, Uromyses are important allergens. Dust mites D. farinae, D.pteronyssinus are also important source of inhalant allergens particularly in the coastal areas of the country. Cockroaches, beetles, weevils, mosquitoes, house flies also contribute towards the aeroallergen load and are allergenically implicated. Avoidance of the indoor and outdoor aeroallergens is recommended for better management of respiratory allergy. PMID:12003301

  4. Investigations of the Host Range of the Corn Cyst Nematode, Heterodera zeae, from Maryland

    PubMed Central

    Ringer, Chloe E.; Sardanelli, Sandra; Krusberg, Lorin R.

    1987-01-01

    The host range of the corn cyst nematode, Heterodera zeae, recently detected in Maryland, was investigated. A total of 269 plant entries, representing 68 families, 172 genera, and 204 species, was inoculated with cysts or a mixture of eggs and second-stage juveniles of H. zeae. The host range of the Maryland population of H. zeae was limited to plants of the Gramineae and included 11 tribes, 33 genera, 42 species, and 77 entries. All 22 corn (Zea mays) cultivars tested were hosts. Other economic hosts included certain cultivars of barley (Hordeum vulgare), oat (Arena sativa), rice (Oryza sativa), sorghum (Sorghum bicolor), sugar cane (Saccharum interspecific hybrid), and wheat (Triticum aestivum). Fall panicum (Panicum dichotomiflorum), a weed species common to cultivated fields in Maryland, was also a host for H. zeae. Other hosts included meadow foxtail (Alopecurus pratensis), Calamagrostis eipgeios, Job's tears (Coix Lachryma-Jobi), green sprangletop (Leptochloa dubia), witchgrass (Panicum capillare), broomcorn (Panicum miliaceum), fountain grass (Pennisetum rueppeli), reed canary grass (Phalaris arundinacea), common reed (Phragmites australis), eastern gamagrass (Tripsacum dactyloides), corn (Zea mays), and teosinte (Zea mexicana). PMID:19290286

  5. Reproductive biology of the biofuel plant Jatropha curcas in its center of origin.

    PubMed

    Rincón-Rabanales, Manuel; Vargas-López, Laura I; Adriano-Anaya, Lourdes; Vázquez-Ovando, Alfredo; Salvador-Figueroa, Miguel; Ovando-Medina, Isidro

    2016-01-01

    In this work, we studied the main characteristics of flowering, reproductive system and diversity of pollinators for the biofuel plant Jatropha curcas (L.) in a site of tropical southeastern Mexico, within its center of origin. The plants were monoecious with inflorescences of unisexual flowers. The male flowers produced from 3062-5016 pollen grains (266-647 per anther). The plants produced fruits with both geitonogamy and xenogamy, although insect pollination significantly increased the number and quality of fruits. A high diversity of flower visiting insects (36 species) was found, of which nine were classified as efficient pollinators. The native stingless bees Scaptotrigona mexicana (Guérin-Meneville) and Trigona (Tetragonisca) angustula (Latreille) were the most frequent visitors and their presence coincided with the hours when the stigma was receptive. It is noteworthy that the female flowers open before the male flowers, favoring xenogamy, which may explain the high genetic variability reported in J. curcas for this region of the world. PMID:26989640

  6. Kinetoplast DNA minicircles: regions of extensive sequence divergence.

    PubMed Central

    Rogers, W O; Wirth, D F

    1987-01-01

    Previous work has shown that the kinetoplast minicircle DNA of Leishmania species exhibits species-specific sequence divergence and this observation has led to the development of a DNA probe-based diagnostic test for leishmaniasis. In the work reported here, we demonstrate that the minicircle is composed of three types of DNA sequences with differing specificities reflecting different rates of DNA sequence change. A library of cloned fragments of kinetoplast DNA (kDNA) from Leishmania mexicana amazonensis was prepared and the cloned subfragments were found to contain DNA sequences with different taxonomic specificities based on hybridization analysis with various species of Leishmania. Four groups of subfragments were found, those that hybridized with a large number of Leishmania sp. as well as sequences unique to the species, subspecies, or isolate. Analysis of nested deletions of a single, full-length minicircle demonstrates that these different taxonomic specificities are contained within a single minicircle. This implies that different regions of a single minicircle have DNA sequences that diverge at different rates. These sequences represent potentially valuable tools in diagnostic, epidemiologic, and ecological studies of leishmaniasis and provide the basis for a model of kDNA sequence evolution. Images PMID:3025880

  7. Cobertura de los sistemas de pensiones y factores asociados al acceso a una pensión de jubilación en México

    PubMed Central

    Murillo-López, Sandra; Venegas-Martínez, Francisco

    2016-01-01

    Resumen Objetivos: obtener estimaciones de indicadores de cobertura de las pensiones por jubilación o retiro para la población mexicana de 65 y más años, y evaluar el impacto que tienen los sistemas de pensiones en las transiciones al retiro de los adultos en edades medias y avanzadas en México. Para ello se utilizan datos microeconómicos provenientes de la Encuesta Nacional de Salud y Envejecimiento. Mediante análisis econométrico se identifican los factores sociodemográficos, económicos, laborales e institucionales que están asociados al acceso a una pensión de jubilación, o bien, a la dependencia de otras fuentes de ingresos. Se encontró que, en México, las transiciones al retiro del mercado de trabajo en las etapas avanzadas del ciclo de vida son limitadas debido a las características eminentemente contributivas de los esquemas de pensiones, los cuales favorecen a la población con trayectorias laborales formales y más estables asociadas a: características de género, oportunidades educativas y posibilidades de inserción en el mercado laboral. PMID:27524936

  8. A case of mucocutaneous leishmaniasis diagnosed by serology.

    PubMed

    Okumura, Yoko; Yamauchi, Asako; Nagano, Isao; Itoh, Makoto; Hagiwara, Keisuke; Takahashi, Kenzo; Uezato, Hiroshi; Maeda, Manabu; Seishima, Mariko

    2014-08-01

    A 43-year-old Japanese Bolivian male had been suffering from a right leg ulcer after an insect bite during his residence in Bolivia. The ulcer healed after herbal medicine treatment. Ten years later, the patient had symptoms of nasal obstruction, nasal bleeding, and pharyngodynia, which were accompanied by a destructive ulcer with surrounding erythema involving the right nostril apex and columella. Papillary, irregular mucosal lesions were seen on the soft palate. Giemsa staining and polymerase chain reaction (PCR) using biopsy specimens of the papillary mucosal lesions on the soft palate failed to identify Leishmania parasites. However, the IgG antibody test was positive for Leishmania (Leishmania) donovani, and the dot enzyme-linked immunosorbent assay (dot-ELISA) using five Leishmania antigens L. (L.) mexicana, L. (L.) amazonensis, Leishmania (Viannia) guyanensis, L. (V.) braziliensis, and L. (V.) panamensis was positive. Combined, the findings suggested mucocutaneous leishmaniasis. Treatment with liposomal amphotericin B was started but was soon terminated because of palpitation, epigastralgia and facial flushing. It is sometimes difficult to identify the parasites in lesions of mucocutaneous leishmaniasis and serological tests are useful for such occasions. PMID:25041360

  9. Meiotic drive of chromosomal knobs reshaped the maize genome.

    PubMed Central

    Buckler, E S; Phelps-Durr, T L; Buckler, C S; Dawe, R K; Doebley, J F; Holtsford, T P

    1999-01-01

    Meiotic drive is the subversion of meiosis so that particular genes are preferentially transmitted to the progeny. Meiotic drive generally causes the preferential segregation of small regions of the genome; however, in maize we propose that meiotic drive is responsible for the evolution of large repetitive DNA arrays on all chromosomes. A maize meiotic drive locus found on an uncommon form of chromosome 10 [abnormal 10 (Ab10)] may be largely responsible for the evolution of heterochromatic chromosomal knobs, which can confer meiotic drive potential to every maize chromosome. Simulations were used to illustrate the dynamics of this meiotic drive model and suggest knobs might be deleterious in the absence of Ab10. Chromosomal knob data from maize's wild relatives (Zea mays ssp. parviglumis and mexicana) and phylogenetic comparisons demonstrated that the evolution of knob size, frequency, and chromosomal position agreed with the meiotic drive hypothesis. Knob chromosomal position was incompatible with the hypothesis that knob repetitive DNA is neutral or slightly deleterious to the genome. We also show that environmental factors and transposition may play a role in the evolution of knobs. Because knobs occur at multiple locations on all maize chromosomes, the combined effects of meiotic drive and genetic linkage may have reshaped genetic diversity throughout the maize genome in response to the presence of Ab10. Meiotic drive may be a major force of genome evolution, allowing revolutionary changes in genome structure and diversity over short evolutionary periods. PMID:10471723

  10. Purification and Partial characterization of Trypanosoma cruzi triosephosphate isomerase.

    PubMed

    Bourguignon, S C; Meirelles, M N; Pacheco, R S; De Simone, S G

    1998-01-01

    The enzyme triosephosphate isomerase (TPI, EC 5.3.1.1) was purified from extracts of epimastigote forms of Trypanosoma cruzi. The purification steps included: hydrophobic interaction chromatography on phenyl-Sepharose, CM-Sepharose, and high performance liquid gel filtration chromatography. The CM-Sepharose material contained two bands (27 and 25 kDa) with similar isoelectric points (pI 9.3-9.5) which could be separated by gel filtration in high performance liquid chromatography. Polyclonal antibodies raised against the porcine TPI detected one single polypeptide on western blot with a molecular weight (27 kDa) identical to that purified from T. cruzi. These antibodies also recognized only one band of identical molecular weight in western blots of several other trypanosomatids (Blastocrithidia culicis, Crithidia desouzai, Phytomonas serpens, Herpertomonas samuelpessoai). The presence of only one enzymatic form of TPI in T. cruzi epimastigotes was confirmed by agarose gel activity assay and its localization was established by immunocytochemical analysis. The T. cruzi purified TPI (as well as other trypanosomatid' TPIs) is a dimeric protein, composed of two identical subunits with an approximate mw of 27,000 and it is resolved on two dimensional gel electrophoresis with a pI of 9.3. Sequence analysis of the N-terminal portion of the 27 kDa protein revealed a high homology to Leishmania mexicana and T. brucei proteins. PMID:9698898

  11. Winter resource wealth drives delayed dispersal and family-group living in western bluebirds.

    PubMed

    Dickinson, Janis L; McGowan, Andrew

    2005-11-22

    Delayed dispersal, where offspring remain with parents beyond the usual period of dependence, is the typical route leading to formation of kin-based cooperative societies. The prevailing explanations for why offspring stay home are variation in resource wealth, in which offspring of wealthy parents benefit disproportionately by staying home, and nepotism, where the tendency for parents to be less aggressive and share food with offspring makes home a superior place to wait to breed. These hypotheses are not strict alternatives, as only wealthy parents have sufficient resources to share. In western bluebirds, Sialia mexicana, sons usually delay dispersal until after winter, gaining feeding advantages through maternal nepotism in a familial winter group. Experimentally reducing resource wealth (mistletoe) by half on winter territories caused sons to disperse in summer, even though their parents remained on the territory during the winter. Only 8% of sons remained with their parents on mistletoe-removal territories compared to 50% of sons on control territories (t(9,10)=3.33, p<0.005). This study is the first to demonstrate that experimentally reducing wealth of a natural food resource reduces delayed dispersal, facilitating nepotism and family-group living. The results clarify the roles of year-round residency, resource limitation and relative wealth outside the breeding season in facilitating the formation of kin-based cooperative societies. PMID:16243691

  12. Long-term production of bioethanol in repeated-batch fermentation of microalgal biomass using immobilized Saccharomyces cerevisiae.

    PubMed

    El-Dalatony, Marwa M; Kurade, Mayur B; Abou-Shanab, Reda A I; Kim, Hoo; Salama, El-Sayed; Jeon, Byong-Hun

    2016-11-01

    Separate hydrolysis fermentation (SHF) and simultaneous saccharification fermentation (SSF) processes were studied for bioethanol production from microalgal biomass. SSF was selected as an efficient process to enhance the bioethanol yield through repeated-batches using immobilized yeast cells. Combined sonication and enzymatic hydrolysis of Chlamydomonas mexicana generated 10.5 and 8.48g/L of ethanol in SSF and SHF, respectively. Yeast utilized maximum portion of total reducing sugar (TRS) reaching a consumption efficiency of 91-98%. A bioethanol yield of 0.5g/g (88.2% of theoretical yield) and volumetric productivity of 0.22g/L/h was obtained after 48h of SSF. Immobilized yeast cells enabled repetitive production of ethanol for 7 cycles displaying a fermentation efficiency up to 79% for five consecutive cycles. The maximum ethanol production was 9.7g/L in 2nd-4th cycles. A total energy recovery of 85.81% was achieved from microalgal biomass in the form of bioethanol. Repeated-batch SSF demonstrated the possibility of cost-effective bioethanol production. PMID:27479800

  13. The Effects of Site Conditions and Mitigation Practices on Success of Establishing the Valley Elderberry Longhorn Beetle and Its Host Plant, Blue Elderberry

    NASA Astrophysics Data System (ADS)

    Holyoak, Marcel; Koch-Munz, Molly

    2008-09-01

    This study performed the first systematic evaluation of the success of habitat mitigation at establishing the threatened Valley elderberry longhorn beetle ( Desmocerus californicus dimorphus) and its host plant, blue elderberry ( Sambucus mexicana). Habitat mitigation performed through enforcement of the U.S. Endangered Species Act represents a tightly controlled form of habitat restoration, facilitating the evaluation of restoration practice. Restoration plantings of blue elderberry have been substantial in our study area, the Central Valley of California. Surveys of 30 mitigation sites and 16 nearby natural sites showed that mitigation sites were a fraction of the size of natural habitat areas (mean = 24%) and contained smaller shrubs. The beetle colonized 53% of mitigation sites and its populations were denser in sites with moderate levels of dead stems on elderberry shrubs, and moderate damage to elderberry stems and bark. This likely indicates that the beetle responds to stressed shrubs, which are likely to contain elevated levels of nitrogen. Beetle density also increased with the size and age of mitigation sites. This indicates a need to make restoration sites as large as possible and to monitor these sites for longer than current guidelines suggest, thereby allowing more time for convergence of natural and mitigation sites. Few factors examined here directly influenced the growth of elderberry shrubs, but elderberry grew more rapidly in sites closer to riparian areas, indicating that such sites should be favored for mitigation sites.

  14. Morphological and physiological comparison of taxa comprising the Sporothrix schenckii complex*

    PubMed Central

    ZHAO, Ming-dan; ZHOU, Xun; LIU, Ting-ting; YANG, Zhi-bang

    2015-01-01

    Based on recent molecular data, it has been suggested that Sporothrix globosa is the main causal agent of sporotrichosis in China. The objective of this study was to compare the morphology, growth characteristics, patterns of carbon source usage, and susceptibility to antifungal agents among Sporothrix strains. A total of 15 clinical strains confirmed to be S. globosa, from three different regions of China, and 11 ex-type strains from the CBS-KNAW biodiversity center were obtained. The elongated conidia of S. pallida, S. variecibatus, S. schenckii, and S. schenckii luriei were clearly different from the subglobose and globose conidia of S. globosa strains. S. schenckii is able to assimilate sucrose, raffinose, and ribitol. Susceptibility profiles of these Sporothrix species were evaluated by measuring minimum inhibitory concentrations (MICs). Fluconazole, itraconazole, terbinafine, and amphotericin B showed good activity against most S. globosa clinical isolates from China. Potassium iodide also showed a low MIC against S. pallida, while fluconazole showed a high MIC for S. mexicana, S. humicola, S. globosa, S. schenckii, and S. inflata; these strains might be considered tolerant. The species showed differences in susceptibility to antifungal drugs and should therefore be properly identified during diagnosis prior to designing therapeutic strategies. PMID:26537212

  15. Construction and analysis of the cDNA subtraction library of yeast and mycelial phases of Sporothrix globosa isolated in China: identification of differentially expressed genes*

    PubMed Central

    Hu, Qing-bi; He, Yu; Zhou, Xun

    2015-01-01

    Species included in the Sporothrix schenckii complex are temperature-dependent with dimorphic growth and cause sporotrichosis that is characterized by chronic and fatal lymphocutaneous lesions. The putative species included in the Sporothrix complex are S. brasiliensis, S. globosa, S. mexicana, S. pallida, S. schenckii, and S. lurei. S. globosa is the causal agent of sporotrichosis in China, and its pathogenicity appears to be closely related to the dimorphic transition, i.e. from the mycelial to the yeast phase, it adapts to changing environmental conditions. To determine the molecular mechanisms of the switching process that mediates the dimorphic transition of S. globosa, suppression subtractive hybridization (SSH) was used to prepare a complementary DNA (cDNA) subtraction library from the yeast and mycelial phases. Bioinformatics analysis was performed to profile the relationship between differently expressed genes and the dimorphic transition. Two genes that were expressed at higher levels by the yeast form were selected, and their differential expression levels were verified using a quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR). It is believed that these differently expressed genes are involved in the pathogenesis of S. globosa infection in China. PMID:26642182

  16. Morphological and physiological comparison of taxa comprising the Sporothrix schenckii complex.

    PubMed

    Zhao, Ming-dan; Zhou, Xun; Liu, Ting-ting; Yang, Zhi-bang

    2015-11-01

    Based on recent molecular data, it has been suggested that Sporothrix globosa is the main causal agent of sporotrichosis in China. The objective of this study was to compare the morphology, growth characteristics, patterns of carbon source usage, and susceptibility to antifungal agents among Sporothrix strains. A total of 15 clinical strains confirmed to be S. globosa, from three different regions of China, and 11 ex-type strains from the CBS-KNAW biodiversity center were obtained. The elongated conidia of S. pallida, S. variecibatus, S. schenckii, and S. schenckii luriei were clearly different from the subglobose and globose conidia of S. globosa strains. S. schenckii is able to assimilate sucrose, raffinose, and ribitol. Susceptibility profiles of these Sporothrix species were evaluated by measuring minimum inhibitory concentrations (MICs). Fluconazole, itraconazole, terbinafine, and amphotericin B showed good activity against most S. globosa clinical isolates from China. Potassium iodide also showed a low MIC against S. pallida, while fluconazole showed a high MIC for S. mexicana, S. humicola, S. globosa, S. schenckii, and S. inflata; these strains might be considered tolerant. The species showed differences in susceptibility to antifungal drugs and should therefore be properly identified during diagnosis prior to designing therapeutic strategies. PMID:26537212

  17. Construction and analysis of the cDNA subtraction library of yeast and mycelial phases of Sporothrix globosa isolated in China: identification of differentially expressed genes.

    PubMed

    Hu, Qing-bi; He, Yu; Zhou, Xun

    2015-12-01

    Species included in the Sporothrix schenckii complex are temperature-dependent with dimorphic growth and cause sporotrichosis that is characterized by chronic and fatal lymphocutaneous lesions. The putative species included in the Sporothrix complex are S. brasiliensis, S. globosa, S. mexicana, S. pallida, S. schenckii, and S. lurei. S. globosa is the causal agent of sporotrichosis in China, and its pathogenicity appears to be closely related to the dimorphic transition, i.e. from the mycelial to the yeast phase, it adapts to changing environmental conditions. To determine the molecular mechanisms of the switching process that mediates the dimorphic transition of S. globosa, suppression subtractive hybridization (SSH) was used to prepare a complementary DNA (cDNA) subtraction library from the yeast and mycelial phases. Bioinformatics analysis was performed to profile the relationship between differently expressed genes and the dimorphic transition. Two genes that were expressed at higher levels by the yeast form were selected, and their differential expression levels were verified using a quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR). It is believed that these differently expressed genes are involved in the pathogenesis of S. globosa infection in China. PMID:26642182

  18. Large scale patterns of abundance and distribution of parasites in Mexican bumblebees.

    PubMed

    Gallot-Lavallée, Marie; Schmid-Hempel, Regula; Vandame, Rémy; Vergara, Carlos H; Schmid-Hempel, Paul

    2016-01-01

    Bumblebees are highly valued for their pollination services in natural ecosystems as well as for agricultural crops. These precious pollinators are known to be declining worldwide, and one major factor contributing to this decline are infections by parasites. Knowledge about parasites in wild bumblebee populations is thus of paramount importance for conservation purposes. We here report the geographical distribution of Crithidia and Nosema, two common parasites of bumblebees, in a yet poorly investigated country: Mexico. Based on sequence divergence of the Cytochrome b and Glycosomal glyceraldehyde phosphate deshydrogenase (gGPDAH) genes, we discovered the presence of a new Crithidia species, which is mainly distributed in the southern half of the country. It is placed by Bayesian inference as a sister species to C. bombi. We suggest the name Crithidia mexicana for this newly discovered organism. A population of C. expoeki was encountered concentrated on the flanks of the dormant volcanic mountain, Iztaccihuatl, and microsatellite data showed evidence of a bottleneck in this population. This study is the first to provide a large-scale insight into the health status of endemic bumblebees in Mexico, based on a large sample size (n=3,285 bees examined) over a variety of host species and habitats. PMID:26678506

  19. Vasoactive and antioxidant activities of plants used in Mexican traditional medicine for the treatment of cardiovascular diseases.

    PubMed

    Ibarra-Alvarado, C; Rojas, A; Mendoza, S; Bah, M; Gutiérrez, D M; Hernández-Sandoval, L; Martínez, M

    2010-07-01

    This study demonstrated that the aqueous extracts of plants employed in Mexican traditional medicine for the treatment of cardiovascular diseases are able to modify the tone of arterial smooth muscle. Agastache mexicana (Kunth) Lint & Epling (Labiatae), Chenopodium murale L. (Chenopodiaceae), Chirantodendron pentadactylon Larreat (Sterculiaceae), Dracocephalum moldavica L. (Labiatae), Psittacanthus calyculatus G. Don (Loranthaceae), Prunus serotina ssp. capuli (Cav. ex Spreng) McVaugh (Rosaceae), and Sechium edule Sw. (Cucurbitaceae) contain secondary metabolites that promote vascular relaxation and display antioxidant activities. As expected, their antioxidant effects showed a significant correlation with the polyphenolics content. However, a lower correlation was found between the antioxidant activity and the maximum vasodilatory effect, suggesting that the vasodilatation elicited by the plant extracts could be only partly attributed to their antioxidant properties. The extract of P. calyculatus, which displayed a maximum vasorelaxant effect that was higher than that of acetylcholine, induced endothelium-dependent vasodilatation. Futhermore, the vasorelaxant response to the P. calyculatus extract was reduced after adding an inhibitor of soluble guanylate cyclase activity, providing evidence that the NO/cGMP pathway is involved. On the other hand, the extracts of Bocconia frutescens L. (Papaveraceae), Magnolia grandiflora L. (Magnoliaceae), and Solanum rostratum Dunal (Solanaceae) induced concentration-dependent contraction of rat aortic rings, suggesting that these plants have potential health benefits for the treatment of ailments such as venous insufficiency. The pharmacological activities of the extracts studied provide scientific support for their ethnomedical use. PMID:20645769

  20. Evaluation of the flora of northern Mexico for in vitro antimicrobial and antituberculosis activity.

    PubMed

    Molina-Salinas, G M; Pérez-López, A; Becerril-Montes, P; Salazar-Aranda, R; Said-Fernández, S; de Torres, N Waksman

    2007-02-12

    The aim of the present study was to evaluate the potential antimicrobial activity of 14 plants used in northeast México for the treatment of respiratory diseases, against drug-sensitive and drug-resistant strains of Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae type b and Mycobacterium tuberculosis. Forty-eight organic and aqueous extracts were tested against these bacterial strains using a broth microdilution test. No aqueous extracts showed antimicrobial activity, whereas most of the organic extracts presented antimicrobial activity against at least one of the drug-resistant microorganisms tested. Methanol-based extracts from the roots and leaves of Leucophyllum frutescens and ethyl ether extract from the roots of Chrysanctinia mexicana showed the greatest antimicrobial activity against the drug-resistant strain of Mycobacterium tuberculosis; the minimal inhibitory concentration (MIC) were 62.5, 125 and 62.5 microg/mL, respectively; methanol-based extract from the leaves of Cordia boissieri showed the best antimicrobial activity against the drug-resistant strain of Staphylococcus aureus (MIC 250 microg/mL); the hexane-based extract from the fruits of Schinus molle showed considerable antimicrobial activity against the drug-resistant strain of Streptococcus pneumoniae (MIC 62.5 microg/mL). This study supports that selecting plants by ethnobotanical criteria enhances the possibility of finding species with activity against resistant microorganisms. PMID:17000069

  1. [Yesterdays and happenings at hospitals of Tehuacán, Puebla, México].

    PubMed

    Fajardo-Ortiz, Guillermo

    2004-01-01

    The history of the hospital of Tehuacán, the second city importance in Puebla State, began in 1744, when the Hospital de San Juan de Dios (Hospital of St. John) was founded; in 1820, it was converted into the Municipal of Civil Hospital. The medical establishement continues to provide services to the poor to this day annual also covers health public workers of services for the municipality. The Hospital de la Cruz Roja Mexicana (The Mexican Red Cross Hospital) was created in 1934 provide medical services for accident victims: The hospital services of the Mexican Institute of Social Security (IMSS) began in 1953; an earthquake destroyed the building in 1973, and a new hospital was inaugurated in 1979; at present, it readers services to 98,000 persons. The Sanatorio del Sagrado Corazón (Sacred Heart Sanatorium) is a private religious institution founded at the beginning of the second half the 20th century. In 1996, the Hospital General (General Hospital) began to provide services, takes care of the poor of the geographical region, and is financed with federal funds. The Instituto de Seguridad y Servicios Sociales para Trabajadores del Estado (Institute of Securtiy and Social Services for the Workers of the State, ISSSTE) has a small hospital with 10 beds, it was founded in 1995. PMID:15087055

  2. Three new species of Ammonicera from the Eastern Pacific coast of North America, with redescriptions and comments on other species of Omalogyridae (Gastropoda, Heterobranchia).

    PubMed

    Sartori, André F; Bieler, Rüdiger

    2014-01-01

    The family Omalogyridae comprises some of the smallest known marine snails. Like all micromolluscs, they have been historically neglected and are underrepresented in faunistic surveys. Based on a few focused studies of the family, 15 valid omalogyrid species were previously recognised in the Indian and Pacific Oceans. To these, we add 3 new species based on a morphological analysis of material in the dry collection of the Natural History Museum of Los Angeles County, applying light and scanning electron microscopies. The new species, Ammonicera mcleani, A. mexicana and A. sleursi, are the first omalogyrid species described from the Eastern Pacific coast of North America. Redescriptions of the Australian omalogyrids Ammonicera sucina (Laseron, 1954) and Omalogyra liliputia (Laseron, 1954) are also presented, detailing for the first time ultrastructural aspects of their shell morphology. Additionally, we present here the first record of Ammonicera binodosa Sleurs, 1985b in Sri Lanka, introduce the new combination Ammonicera vangoethemi (Sleurs, 1985c) for Omalogyra vangoethemi, and formally remove Transomalogyra Palazzi & Gaglini, 1979 from Omalogyridae by fixing its type species as Homalogyra densicostata Jeffreys, 1884. Finally, we present lists, geographic records and a bibliography of all currently recognised omalogyrid species in the Indian, Pacific and Antarctic Oceans, as well as an update to a previously published compilation of the Atlantic and Mediterranean representatives of the family. PMID:25544067

  3. Taxonomy, diversity, temporal and geographical distribution of Cutaneous Leishmaniasis in Colombia: A retrospective study

    PubMed Central

    Ramírez, Juan David; Hernández, Carolina; León, Cielo M.; Ayala, Martha S.; Flórez, Carolina; González, Camila

    2016-01-01

    Leishmaniases are tropical zoonotic diseases, caused by kinetoplastid parasites from the genus Leishmania. New World (NW) species are related to sylvatic cycles although urbanization processes have been reported in some South American Countries such as Colombia. Currently, few studies show the relative distribution of Leishmania species related to cutaneous Leishmaniasis (CL) in South America due to the lack of accurate surveillance and public health systems. Herein, we conducted a systematic estimation of the Leishmania species causing CL in Colombia from 1980 to 2001 via molecular typing and isoenzymes. A total of 327 Leishmania isolates from humans, sandflies and reservoirs were typed as L. panamensis 61.3% (201), L. braziliensis 27.1% (88), L. lainsoni 0.6% (2), L. guyanensis 0.9% (3), L. infantum chagasi 4% (12), L. equatoriensis 0.6% (2), L. mexicana 2.1% (8), L. amazonensis 2.8% (9) and L. colombiensis 0.6% (2). This is the first report of two new Leishmania species circulating in Colombia and suggests the need to convince the Colombian government about the need to deploy and standardize tools for the species identification to provide adequate management to individuals suffering this pathology. PMID:27328969

  4. Reproductive biology of the biofuel plant Jatropha curcas in its center of origin

    PubMed Central

    Rincón-Rabanales, Manuel; Vargas-López, Laura I.; Adriano-Anaya, Lourdes; Salvador-Figueroa, Miguel

    2016-01-01

    In this work, we studied the main characteristics of flowering, reproductive system and diversity of pollinators for the biofuel plant Jatropha curcas (L.) in a site of tropical southeastern Mexico, within its center of origin. The plants were monoecious with inflorescences of unisexual flowers. The male flowers produced from 3062–5016 pollen grains (266–647 per anther). The plants produced fruits with both geitonogamy and xenogamy, although insect pollination significantly increased the number and quality of fruits. A high diversity of flower visiting insects (36 species) was found, of which nine were classified as efficient pollinators. The native stingless bees Scaptotrigona mexicana (Guérin-Meneville) and Trigona (Tetragonisca) angustula (Latreille) were the most frequent visitors and their presence coincided with the hours when the stigma was receptive. It is noteworthy that the female flowers open before the male flowers, favoring xenogamy, which may explain the high genetic variability reported in J. curcas for this region of the world. PMID:26989640

  5. Reactivation of triosephosphate isomerase from three trypanosomatids and human: effect of suramin.

    PubMed Central

    Gao, X G; Garza-Ramos, G; Saavedra-Lira, E; Cabrera, N; De Gómez-Puyou, M T; Perez-Montfort, R; Gómez-Puyou, A

    1998-01-01

    The reactivation of the homodimeric triosephosphate isomerases (TIMs) from Trypanosoma brucei, T. cruzi, Leishmania mexicana and humans was determined after their denaturation with guanidine hydrochloride. In the range of 2-32 microg of T. brucei TIM per ml and 0.2-5 microg of the other enzymes per ml, the rate and extent of TIM reactivation depended on protein concentration, indicating that at these protein concentrations, the rate-limiting step of reactivation is monomer association and not monomer folding. The rate of monomer association was more than one order of magnitude lower in the T. brucei enzyme than in the other three enzymes. Suramin is a drug of choice in the treatment of sleeping sickness, but its mechanism of action is not known. At micromolar concentrations, Suramin inhibited the reactivation of the four enzymes, but the extent of inhibition by Suramin decreased with increasing protein concentration as consequence of a diminution of the life time of the folded monomer. Since the life time of the monomer of T. brucei TIM is longer than that of the other enzymes, Suramin is a more effective inhibitor of the reactivation of TIM from T. brucei, particularly at monomer concentrations above 1 microg of protein per ml (monomer concentration approx. 37 nM). Compounds that are structurally related to Suramin also inhibit TIM reactivation; their effect was about five times more pronounced in the enzyme from T. brucei than in human TIM. PMID:9576855

  6. Female bias for enlarged male body and dorsal fins in Xiphophorus variatus.

    PubMed

    MacLaren, R David; Gagnon, John; He, Ran

    2011-06-01

    Female preference for male fin elaborations in Poeciliid fishes may be driven by a sensory bias for increased lateral projection area (LPA) that has existed since the lineages diverged from a common ancestor. Previous research supports this hypothesis demonstrating female Poecilia latipinna, Poecilia mexicana, and Poecilia reticulata prefer males of larger body and dorsal fin size, but exhibit no such preferences when controlling for total LPA. In the current study, we further tested this hypothesis by presenting female platys, Xiphophorus variatus, with pairs of dummy males differing in: (1) body size (holding dorsal fin size constant); (2) dorsal fin size (holding body size constant); and (3) dorsal fin: body size ratio (holding total LPA constant). Females spent more time near dummies of greater body and dorsal fin size; however, in the third experiment, neither fin size, body size, nor any particular dorsal fin+body size combination was preferred. These results provide additional support for the LPA and sensory bias hypotheses, demonstrating that female X. variatus not only prefer males with "swords", but sailfin-like dorsal fins as well when body size is held constant. Shared preference for increased LPA is consistent with common ancestry of the sensory/neural systems in females of all four species. PMID:21457765

  7. Report on the Threatened Valley Elderberry Longhorn Beetle and its Elderberry Food Plant at the Lawrence Livermore National Laboratory--Site 300

    SciTech Connect

    Arnold, Ph.D., R A; Woollett, J

    2004-11-16

    This report describes the results of an entomological survey in 2002 to determine the presence of the federally-listed, threatened Valley Elderberry Longhorn Beetle or ''VELB'' (Desmocerus culifornicus dimorphus: Coleoptera, Cerambycidae) and its elderberry food plant (Sumbucus mexicana: Caprifoliaceae) on the Lawrence Livermore National Laboratory's (LLNL) Experimental Test Site, known as Site 300. In addition, an area located immediately southeast of Site 300, which is owned and managed by the California Department of Fish and Game (CDFG), but secured by LLNL, was also included in this survey. This report will refer to the survey areas as the LLNL-Site 300 and the CDFG site. The 2002 survey included mapping the locations of elderberry plants that were observed using a global positioning system (GPS) to obtain positional coordinates for every elderberry plant at Site 300. In addition, observations of VELB adults and signs of their infestation on elderberry plants were also mapped using GPS technology. LLNL requested information on the VELB and its elderberry food plants to update earlier information that had been collected in 1991 (Arnold 1991) as part of the 1992 EIS/EIR for continued operation of LLNL. No VELB adults were observed as part of this prior survey. The findings of the 2002 survey reported herein will be used by LLNL as it updates the expected 2004 Environmental Impact Statement for ongoing operations at LLNL, including Site 300.

  8. Microbial populations related to PAH biodegradation in an aged biostimulated creosote-contaminated soil.

    PubMed

    Lladó, Salvador; Jiménez, Nuria; Viñas, Marc; Solanas, Anna Maria

    2009-09-01

    A previous bioremediation survey on a creosote-contaminated soil showed that aeration and optimal humidity promoted depletion of three-ringed polycyclic aromatic hydrocarbons (PAHs), but residual concentrations of four-ringed benzo(a)anthracene (B(a)A) and chrysene (Chry) remained. In order to explain the lack of further degradation of heavier PAHs such as four-ringed PAHs and to analyze the microbial population responsible for PAH biodegradation, a chemical and microbial molecular approach was used. Using a slurry incubation strategy, soil in liquid mineral medium with and without additional B(a)A and Chry was found to contain a powerful PAH-degrading microbial community that eliminated 89% and 53% of the added B(a)A and Chry, respectively. It is hypothesized that the lack of PAH bioavailability hampered their further biodegradation in the unspiked soil. According to the results of the culture-dependent and independent techniques Mycobacterium parmense, Pseudomonas mexicana, and Sphingobacterials group could control B(a)A and Chry degradation in combination with several microorganisms with secondary metabolic activity. PMID:19153811

  9. Review of the genus Arhaphe (Hemiptera: Heteroptera: Largidae) with descriptions of nine new species from Central America.

    PubMed

    Stehlík, Jaroslav L; Brailovsky, Harry

    2016-01-01

    Nine new species of the genus Arhaphe Herrich-Schaeffer, 1850 are described: A. ferruginea sp. nov. from Mexico (Guerrero), A. flavoantennata sp. nov. from Costa Rica (Guanacaste Province), Honduras (Intibuca Department) and Nicaragua (Granada Province), A. hirsuta sp. nov. from Mexico (Oaxaca), A. kmenti sp. nov. from Mexico (Guanajuato, Michoacán), A. longula sp. nov. from Mexico (Guerrero), A. magna sp. nov. from Mexico (Colima), A. myrmicoides sp. nov. from Mexico (Guerrero, Nayarit), A. oaxacana sp. nov. from Mexico (Oaxaca), and A. pilifera sp. nov. from Mexico (Nayarit). The fifth instar larvae of A. arguta (Bliven, 1956), A. carolina Herrich-Schaeffer, 1850, A. cicindeloides Walker, 1873, A. flavoantennata sp. nov., A. mexicana Halstead, 1972, and A. morelensis Brailovsky & Marquez, 1974 are described. Additional state faunistic records of the already described species are provided: A. arguta (Bliven, 1956) (Mexico: Guanajuato, Sonora), A. capitata Halstead, 1972 (Costa Rica; Mexico: Chiapas, Estado de México, Guerrero, Michoacán, Oaxaca, Puebla), A. carolina Herrich-Schaeffer, 1850 (Mexico: Guanajuato, Tamaulipas), A. cicindeloides Walker, 1873 (Mexico: Hidalgo, Querétaro), A. deviatica Brailovsky, 1981 (Nicaragua), A. furcata Brailovsky, 1981 (Mexico: Michoacán), A. halsteadi Brailovsky, 1981 (Mexico: Oaxaca), A. mimetica Barber, 1911 (Mexico: Chihuahua), A. morelensis Brailovsky & Marquez, 1974 (Mexico: Estado de México, Guerrero, Puebla), and A. rustica Brailovsky, 1981 (Mexico: Morelos, Oaxaca). PMID:27394508

  10. Treatment of cutaneous leishmaniasis with localized current field (radio frequency) in Tabasco, Mexico.

    PubMed

    Velasco-Castrejon, O; Walton, B C; Rivas-Sanchez, B; Garcia, M F; Lazaro, G J; Hobart, O; Roldan, S; Floriani-Verdugo, J; Munguia-Saldana, A; Berzaluce, R

    1997-09-01

    In Mexico cutaneous leishmaniasis (CL) occurs in 17 of 32 States, and is a serious public health problem. This is a report of treatment of CL patients in the State of Tabasco, Mexico with a localized current field-radio frequency (LCF-RF) device to generate precisely controlled heat as an alternative to prohibitively expensive drug treatment. It was not a controlled clinical trial, but rather an evaluation of the feasibility of this form of treatment for all CL patients encountered in the endemic area. A total of 201 previously diagnosed patients with CL caused by Leishmania mexicana were treated with a portable Thermosurgery LCF-RF generator powered by rechargeable batteries. The ages of the patients ranged from two to 75 years; the sex distribution was 63% males and 37% females. A single device was used in five different municipalities. Lesions were first anesthetized with 1% lidocaine HCl and moistened with normal saline solution. Treatment consisted of a single application that produced 50 degrees C for 30 sec. After four weeks, 122 patients were available for evaluation, of which 95% were totally cured; (even those involving ear cartilage, which respond poorly to antimonials). At eight weeks post-treatment, 191 patients were evaluated, with a total cure rate of 90%. This form of treatment proved to be effective and convenient for use in primary health care facilities in Mexico and has many advantages over traditional forms of therapy. PMID:9311641

  11. Seroepidemiological studies of cutaneous leishmaniasis in the Campeche state of México.

    PubMed

    Monroy-Ostria, A; Sosa-Cabrera, T; Rivas-Sanchez, B; Ruiz-Tuyu, R; Mendoza-Gonzalez, A R; Favila-Castillo, L

    1997-01-01

    Seroepidemiological studies of cutaneous leishmaniasis were carried out in 169 individuals in a rural area of the Campeche state of México. Fifty showed cutaneous lesions suggestive of leishmaniasis, 70% were parasite positive and 96% skin test positive. An overall 40% positivity to skin test with Montenegro's antigen was found. Most of the affected individuals were males from 11 to 30 years-old. Antibodies were determined by immunofluorescent antibody test (IFA) and by Western blot. Two antigen preparations were used, one from a Leishmania mexicana strain which produced localized cutaneous leishmaniasis (LCL) and the other from a diffuse cutaneous leishmaniasis (DCL). In the general population from the area of study 19% gave positive IFA tests with DCL antigen and 20% with LCL antigen while for the patients 67% gave positive IFA tests with DCL and 71% with LCL. By Western blot analysis most of the patients recognized more antigens in the DCL than in the LCL strain. In the DCL strain 78% of patients recognized a 105 kDa, 34% a 139 kDa, 28% a 117 kDa and 26% a 205 kDa MW antigen. In the LCL strain 40% of patients recognized a 205 kDa and 22% a 175 kDa antigens. PMID:9302408

  12. Cutaneous leishmaniasis caused by members of Leishmania braziliensis complex in Nayarit, State of Mexico.

    PubMed

    Sanchez-Tejeda, G; Rodríguez, N; Parra, C I; Hernandez-Montes, O; Barker, D C; Monroy-Ostria, A

    2001-01-01

    An epidemiological study was carried out in the northern Mexican state, Nayarit. Fourteen patients with possible cutaneous leishmaniasis skin lesions gave positive Montenegro skin tests. Biopsies were taken from the skin ulcer and analyzed by polymerase chain reaction (PCR) with specific primers for the Leishmania mexicana complex; however all biopsies were not amplified. PCR carried out with specific primers for the L. braziliensis complex resulted in the amplification of all patient DNA. DNA from 12 out of 14 biopsies gave positive amplification with primers species specific for L. (Viannia) braziliensis and hybridized with a species specific L. (V.) braziliensis probe. These results demonstrate the presence in Nayarit of at least two members of the L. braziliensis complex. Most of the cutaneous lesions were caused by L. (V.) braziliensis and two by another species belonging to the L. braziliensis complex. As far as we are aware, this is the first report of L. (V.) braziliensis in Nayarit. The main risk factor associated with the contraction of this disease in Nayarit is attributed to working on coffee plantations. PMID:11285471

  13. Candida heliconiae sp. nov., Candida picinguabensis sp. nov. and Candida saopaulonensis sp. nov., three ascomycetous yeasts from Heliconia velloziana (Heliconiaceae).

    PubMed

    Ruivo, Carla C C; Lachance, Marc-André; Rosa, Carlos A; Bacci, Maurício; Pagnocca, Fernando C

    2006-05-01

    Strains belonging to three novel yeast species, Candida heliconiae (four isolates), Candida picinguabensis (three isolates) and Candida saopaulonensis (two isolates), were recovered in the year 2000 from water of flower bracts of Heliconia velloziana L. Emigd. (Heliconiaceae) found in a forest ecosystem site in an Atlantic rainforest of south-eastern Brazil. C. picinguabensis and C. saopaulonensis were nearly identical in morphology and physiology, but sequence divergence in the D1/D2 domain of the large-subunit rDNA indicated that they should be regarded as different species. They belong to the Metschnikowiaceae clade. C. heliconiae had affinities to Pichia mexicana and related species, but was genetically isolated from all currently accepted species in that group. The type strains are C. heliconiae UNESP 00-91C1T (=CBS 10000T=NRRL Y-27813T), C. picinguabensis UNESP 00-89T (=CBS 9999T=NRRL Y-27814T) and C. saopaulonensis UNESP 00-99T (=CBS 10001T=NRRL Y-27815T). PMID:16627669

  14. The Volcano Adventure Guide

    NASA Astrophysics Data System (ADS)

    Lopes, Rosaly

    2005-02-01

    This guide contains vital information for anyone wishing to visit, explore, and photograph active volcanoes safely and enjoyably. Following an introduction that discusses eruption styles of different types of volcanoes and how to prepare for an exploratory trip that avoids volcanic dangers, the book presents guidelines to visiting 42 different volcanoes around the world. It is filled with practical information that includes tour itineraries, maps, transportation details, and warnings of possible non-volcanic dangers. Three appendices direct the reader to a wealth of further volcano resources in a volume that will fascinate amateur enthusiasts and professional volcanologists alike. Rosaly Lopes is a planetary geology and volcanology specialist at the NASA Jet Propulsion Laboratory in California. In addition to her curatorial and research work, she has lectured extensively in England and Brazil and written numerous popular science articles. She received a Latinas in Science Award from the Comision Feminil Mexicana Nacional in 1991 and since 1992, has been a co-organizer of the United Nations/European Space Agency/The Planetary Society yearly conferences on Basic Science for the Benefit of Developing Countries.

  15. Lilliput effect in a retroplumid crab (Crustacea: Decapoda) across the K/Pg boundary

    NASA Astrophysics Data System (ADS)

    Martínez-Díaz, José Luis; Phillips, George E.; Nyborg, Torrey; Espinosa, Belinda; Távora, Vladimir de Araújo; Centeno-García, Elena; Vega, Francisco J.

    2016-08-01

    The genus Costacopluma (Brachyura: Decapoda: Retroplumidae) had a wide distribution during the early Paleogene and is currently represented by 14 species across the Late Cretaceous and early Paleogene. Described early Paleogene species have a smaller mean body size compared to Campanian-Maastrichtian populations of Africa, northeastern Mexico, and southeastern United States. Originally described from the Paleocene and Eocene of Alabama, Costacopluma grayi Feldmann and Portell, 2007, is now documented from the uppermost Maastrichtian (66.2 Ma) of northeastern Mexico and Mississippi and Lower Paleocene of Arkansas, all representing medium size specimens. The morphological features of latest Maastrichtian (66.2 Ma) individuals are identical to those observed among populations of C. grayi from the Paleogene of Alabama and Arkansas, which have a smaller mean size. This size reduction, or dwarfism, in C. grayi across the K-Pg boundary is an example of the Lilliput effect. Dwarfism has been documented in several invertebrate groups as a response to environmental stress, but this is the first record of the Lilliput effect in brachyuran crustaceans. The stratigraphic and geographic range for Costacopluma mexicana Vega and Perrilliat, 1989, is extended to the upper Campanian in northeastern Mexico and lower Maastrichtian in Mississippi and is suggested as a possible ancestor of C. grayi. Different preservational modes for this species in northeastern Mexico are discussed.

  16. Winter resource wealth drives delayed dispersal and family-group living in western bluebirds

    PubMed Central

    Dickinson, Janis L; McGowan, Andrew

    2005-01-01

    Delayed dispersal, where offspring remain with parents beyond the usual period of dependence, is the typical route leading to formation of kin-based cooperative societies. The prevailing explanations for why offspring stay home are variation in resource wealth, in which offspring of wealthy parents benefit disproportionately by staying home, and nepotism, where the tendency for parents to be less aggressive and share food with offspring makes home a superior place to wait to breed. These hypotheses are not strict alternatives, as only wealthy parents have sufficient resources to share. In western bluebirds, Sialia mexicana, sons usually delay dispersal until after winter, gaining feeding advantages through maternal nepotism in a familial winter group. Experimentally reducing resource wealth (mistletoe) by half on winter territories caused sons to disperse in summer, even though their parents remained on the territory during the winter. Only 8% of sons remained with their parents on mistletoe-removal territories compared to 50% of sons on control territories (t9,10=3.33, p<0.005). This study is the first to demonstrate that experimentally reducing wealth of a natural food resource reduces delayed dispersal, facilitating nepotism and family-group living. The results clarify the roles of year-round residency, resource limitation and relative wealth outside the breeding season in facilitating the formation of kin-based cooperative societies. PMID:16243691

  17. Binding Energy Calculations for Novel Ternary Ionic Lattices

    NASA Astrophysics Data System (ADS)

    Rodríguez-Mijangos, Ricardo; Vazquez-Polo, Gustavo

    2002-03-01

    Theoretical calculations for the binding energy between metalic ions and negative ions on a novel ternary ionic lattice is carried out for several solid solutions prepared with different concentrations and characterized recently (1). The ternary lattices that reach a good miscibility are: KCl(x)KBr(y)RbCl(z) in three different concentrations: (x=y=z=0.33), (x=0.5, y=0.25, z=0.25) and (x=0.33, y=0.07, z=0.60). The binding energy for these novel structures is calculated from the lattice constants obtained by X ray diffractometry analysis performed on the samples and the Vegard law (2). For the repulsive force exponent m, an average of the m values was considered. The energy values obtained by the Born´expression are compared with corresponding energy values from the lattice with more complex expressions, such as the Born Mayer, Born-Van der Walls. There is a good aggreement between all these calculations. (1)R. R. Mijangos, A. Cordero-Borboa, E. Alvarez, M. Cervantes, Physics Letters A 282 (2001) 195-200. (2) G. Vazquez-Polo, R. R. Mijangos et al. Revista Mexicana de Fisica, 47, Diciembre 2001. In Press.

  18. Atmospheric secondary charged cosmic radiation at a place of 11.5 GV geomagnetic cut-off

    NASA Astrophysics Data System (ADS)

    Azcarate, I. N.

    2002-04-01

    An experiment performed with a balloon-borne plastic scintillator is described. The detector system was transported by a stratospheric balloon, that was launched from Reconquista, province of Santa Fe, Argentina, on 24 February 1992. The geomagnetic cut-off of the site was 11.5 GV. The energy-loss spectra of both the atmospheric gamma radiation ( for E^γ>= 4.15 MeV) and the charged component of the secondary cosmic radiation were alternatively measured at different altitudes, during the ascent of the balloon, and at ceiling altitude. The author analyzed the atmospheric gamma-ray spectrum in a previous paper ( Azcárate, 2000). It was necessary to perform the computation of the response of the detector to the charged radiation in order to explain , at least qualitatively, the energy-loss spectrum in the detector produced by this type of radiation. It is argued that at ceiling altitude the observed feature in the spectrum is produced mainly by relativistic muons falling horizontally upon the detector. The growth curve for the counting rate below this feature and the intensity of relativistic μ-mesons were also obtained. References : Azcárate, I.N., Revista Mexicana de Astronomía y Astrofísica, 36, 81, 2000.

  19. New Data in the ADS Abstract and Article Service

    NASA Astrophysics Data System (ADS)

    Eichhorn, G.; Accomazzi, A.; Grant, C. S.; Kurtz, M. J.; Murray, S. S.

    1996-05-01

    In the last few months the data holdings in the ADS have been considerably expanded. In the abstracts databases we have included over 50,000 abstracts from SPIE conference proceedings (provided by SPIE), a complete set of references for lunar and planetary sciences, and abstracts from recent Lunar and Planetary Institute sponsored conferences (both provided by the Lunar and Planetary Institute). We also extended our cooperation with the CDS in Strasbourg, France by providing a link to the list of objects that are in the SIMBAD database for each reference. The ADS article service now holds full-text articles for 20 years of the Astrophysical Journal Letters, the Astronomical Journal, and the Publications of the Astronomical Society of the Pacific, and 5 years of the Astrophysical Journal on-line. The following journals are being processed and some may be on-line for this AAS meeting: Astrophysical Journal (1975-1989), Astronomy and Astrophysics, Proceedings of the Astronomical Society of Australia, Publications of the Astronomical Society of Japan, Revista Mexicana, Bulletin of the Astronomical Society of India, Obs. Reports of Skalnate Pleso, and Baltic Astronomy. We are now working with two scanning companies to speed up the scanning process and hope that by the end of the year we have all of these journals completely on-line for at least the period 1975 - 1995. Usage statistics for Jan - March 1996: Users Queries References Full Abstracts retrieved retrieved ------------------------------------------- 13,823 365,812 7,953,930 309,866

  20. Hantavirus testing in small mammal populations of northcentral New Mexico

    SciTech Connect

    Biggs, J.; Bennett, K.; Foxx, T.

    1995-07-01

    In 1993, an outbreak of a new strain of hantavirus in the southwestern US indicated that deer mice (Peromyscus maniculatus) was the primary carrier of the virus. In 1993 and 1994, the Ecological Studies Team (EST) at Los Alamos National Laboratory surveyed small mammal populations in Los Alamos County, New Mexico, primarily for ecological risk assessment (ecorisk) studies. At the request of the Centers for Disease Control (CDC) and the School of Medicine at the University of New Mexico, EST also collected blood samples from captured animals for use in determining seroprevalence of hantavirus in this region due to the recent outbreak of this virus in the four-comers region of the Southwest. The deer mouse was the most commonly captured species during the tripping sessions. Other species sampled included harvest mice (Reithrodontomys megalotis), least chipmunk (Eutamias minimus), long-tailed vole (Microtus longicaudus), Mexican woodrat (Neotoma mexicana), and brush mouse (Peromyscus boylii). The team collected blood samples from tripped animals following CDC`s suggested guidelines. Results of the 1993 and 1994 hantavirus testing identified a total overall seroprevalence of approximately 5.5% and 4.2%, respectively. The highest seroprevalence rates were found in deer mice seri (3--6%), but results on several species were inconclusive; further studies will be necessary, to quantify seroprevalence rates in those species. Seroprevalence rates for Los Alamos County were much lower than elsewhere in the region.

  1. First evidence for heritable variation in cooperative breeding behaviour.

    PubMed

    Charmantier, Anne; Keyser, Amber J; Promislow, Daniel E L

    2007-07-22

    Understanding the evolution of complex social behaviours, such as cooperative breeding, is a fundamental problem in evolutionary biology, which has attracted much theoretical and empirical interest. Variation within and between species in the frequency of helping behaviour has been typically associated with variation in direct costs and benefits due to ecological constraints, or with indirect fitness payoffs (i.e. kin selection). Here, we provide the first evidence that individual variation in cooperative behaviour within a natural population also has a heritable component. Using a seven-generation pedigree in a wild population of western bluebirds (Sialia mexicana), we show significant heritable variation for the propensity to help rather than breed, as well as for the probability of having a helper at the nest. We also document a strong positive relationship between a bird's lifespan and its prospect of receiving help when breeding, in accordance with earlier comparative studies across species. These findings provide useful insights into the possible mechanisms which have led to the evolution of cooperative breeding and other social systems. PMID:17490945

  2. Phlebotomine sandflies and leishmaniasis risks in Colombian coffee plantations under two systems of cultivation.

    PubMed

    Alexande; Agudelo, L A; Navarro, F; Ruiz, F; Molina, J; Aguilera, G; Quiñones, M L

    2001-12-01

    The phlebotomine sandfly fauna of traditional (shaded) and intensified (unshaded) coffee plantations in Colombia was sampled by a variety of methods and the species composition and density under the two systems compared. Twenty species of Lutzomyia sandflies (Diptera: Psychodidae: Phlebotominae) were collected, of which eight were found only in the 'Coffee Axis' ('Eje Cafetero') of the departments of Caldas, Risaralda and Quindio, six were exclusive to the department of Norte de Santander and six occurred in both regions. Four species were collected only in traditional plantations and two exclusively in intensified ones. At least 13 species occurred in both plantation types. Fifteen species are opportunistic man-biters and eight are suspected vectors of leishmaniasis caused by Le. braziliensis, Le. panamensis or Le. mexicana. Seven species were collected inside houses and may be involved in intradomiciliary transmission of Leishmania. The dominant species in Norte de Santander was Lu. spinicrassa, which made up 93.8% of all the sandflies collected in this department. This species was absent from the Eje Cafetero and a number of others among the 15 recorded there might be responsible for Leishmania transmission in this region, including Lu. trapidoi, Lu. yuilli, Lu. gomezi, L. hartmanni and Lu. ovallesi. Sandfly population densities were significantly higher in traditional plantations than in intensified ones. Residents of traditional plantations were able to describe sandflies in significantly more detail than those of intensified plantations, based on seven basic characteristics related to the appearance and biting behaviour of the insects. PMID:11776455

  3. Criticisms biologically unwarranted and analytically irrelevant: Reply to Rominger et al.

    USGS Publications Warehouse

    Bender, L.C.; Weisenberger, M.E.

    2009-01-01

    The criticisms of Rominger et al. (2008) of our retrospective analysis of desert bighorn sheep (DBS; Ovis canadensis mexicana) dynamics in the San Andres Mountains of south-central New Mexico, USA, contained many biological errors and analytical oversights. Herein, we show that Rominger et al. (2008) 1) overstated both magnitude and potential effect of predator removal; 2) incorrectly claimed that our total precipitation (TP) model did not fit the data when TP correctly classed ???66 of subsequent population increases and declines (P ??? 0.063); 3) presented a necessary prerequisite of the exponential model (serial correlation between Nt and Nt1) as the key relationship in the DBS data, when it merely reflected that DBS are strongly K-selected and was irrelevant to our hypothesis tests specific to factors affecting the instantaneous rate of population increase (r); 4) greatly oversimplified relationships among precipitation, arid environments, and DBS; and 5) advocated a time for collection of lamb/female (L/F) ratio data that was unrelated to any meaningful period in the biological year of DBS and consequently presented L/F ratio data unrelated to observed dynamics of DBS. In contrast, the L/F ratios used in Bender and Weisenberger (2005) correctly predicted annual changes and were correlated with long-term population rates of change.

  4. Cooperation and competition: nepotistic tolerance and intrasexual aggression in western bluebird winter groups

    USGS Publications Warehouse

    Dickinson, J.L.; Euaparadorn, M.; Greenwald, K.; Mitra, C.; Shizuka, D.

    2009-01-01

    Two hypothesized benefits of delayed dispersal are access to resources and prolonged brood care (or??parental nepotism). Resource abundance (mistletoe wealth) is a key factor influencing whether sons stay home in western bluebirds, Sialia mexicana, but nepotism is also observed. Western bluebird sons commonly remain in their family groups throughout the winter, whereas daughters usually disperse before winter. Because pairing often takes place in winter groups, with newly formed pairs settling on exclusive all-purpose territories in spring, selection for sexual competition and nepotism co-occur and may simultaneously influence patterns of aggression within groups. We measured aggression at mealworm feeder stations, finding evidence of (1) intrasexual aggression against unrelated group members by experienced breeders of both sexes and (2) nepotism towards sons and daughters by experienced breeder females but not by experienced breeder males. Females showed much higher levels of aggression towards same-sex immigrants than males did. Experienced breeder males did not evict their sons from the natal territory, but they were 12 times more aggressive towards sons than breeder females were towards daughters. They were also equally aggressive towards sons and immigrant males, suggesting that local breeding competition and the benefits of intrasexual dominance counter the benefits of paternal nepotism towards sons. ?? 2009 The Association for the Study of Animal Behaviour.

  5. Megabase-Scale Inversion Polymorphism in the Wild Ancestor of Maize

    PubMed Central

    Fang, Zhou; Pyhäjärvi, Tanja; Weber, Allison L.; Dawe, R. Kelly; Glaubitz, Jeffrey C.; González, José de Jesus Sánchez; Ross-Ibarra, Claudia; Doebley, John; Morrell, Peter L.; Ross-Ibarra, Jeffrey

    2012-01-01

    Chromosomal inversions are thought to play a special role in local adaptation, through dramatic suppression of recombination, which favors the maintenance of locally adapted alleles. However, relatively few inversions have been characterized in population genomic data. On the basis of single-nucleotide polymorphism (SNP) genotyping across a large panel of Zea mays, we have identified an ∼50-Mb region on the short arm of chromosome 1 where patterns of polymorphism are highly consistent with a polymorphic paracentric inversion that captures >700 genes. Comparison to other taxa in Zea and Tripsacum suggests that the derived, inverted state is present only in the wild Z. mays subspecies parviglumis and mexicana and is completely absent in domesticated maize. Patterns of polymorphism suggest that the inversion is ancient and geographically widespread in parviglumis. Cytological screens find little evidence for inversion loops, suggesting that inversion heterozygotes may suffer few crossover-induced fitness consequences. The inversion polymorphism shows evidence of adaptive evolution, including a strong altitudinal cline, a statistical association with environmental variables and phenotypic traits, and a skewed haplotype frequency spectrum for inverted alleles. PMID:22542971

  6. Cytotoxic effects of bangladeshi medicinal plant extracts.

    PubMed

    Uddin, Shaikh J; Grice, I Darren; Tiralongo, Evelin

    2011-01-01

    To investigate the cytotoxic effect of some Bangladeshi medicinal plant extracts, 16 Bangladeshi medicinal plants were successively extracted with n-hexane, dichloromethane, methanol and water. The methanolic and aqueous extracts were screened for cytotoxic activity against healthy mouse fibroblasts (NIH3T3) and three human cancer-cell lines (gastric: AGS; colon: HT-29; and breast: MDA-MB-435S) using the MTT assay. Two methanolic extracts (Hygrophila auriculata and Hibiscus tiliaceous) and one aqueous extract (Limnophila indica) showed no toxicity against healthy mouse fibroblasts, but selective cytotoxicity against breast cancer cells (IC(50) 1.1-1.6 mg mL(-1)). Seven methanolic extracts from L. indica, Clerodendron inerme, Cynometra ramiflora, Xylocarpus moluccensis, Argemone mexicana, Ammannia baccifera and Acrostichum aureum and four aqueous extracts from Hygrophila auriculata, Bruguiera gymnorrhiza, X. moluccensis and Aegiceras corniculatum showed low toxicity (IC(50) > 2.5 mg mL(-1)) against mouse fibroblasts but selective cytotoxicity (IC(50) 0.2-2.3 mg mL(-1)) against different cancer cell lines. The methanolic extract of Blumea lacera showed the highest cytotoxicity (IC(50) 0.01-0.08 mg mL(-1)) against all tested cell lines among all extracts tested in this study. For some of the plants their traditional use as anticancer treatments correlates with the cytotoxic results, whereas for others so far unknown cytotoxic activities were identified. PMID:19706693

  7. Autoreporte de exposición a publicidad y promoción de tabaco en una cohorte de fumadores mexicanos: Antes y durante la publicación de la Ley General para el Control del tabaco en 2008

    PubMed Central

    Hernández, Rosaura Pérez; Thrasher, James F.; Bolaños, Rosibel Rodríguez; Gutiérrez, Inti Barrientos; Hernández, Norma A Ibañez

    2015-01-01

    Resumen Objetivo Determinar en población fumadora el nivel de exposición a la mercadotecnia por parte de la Industria Tabacalera (IT), a través de diferentes métodos de promocionar sus productos de tabaco, antes y durante la publicación de la Ley General para el Control del Tabaco (LGCT) en 2008. Material y métodos Estudio de cohorte en fumadores adultos (n=941 pre-LGCT y n=1051 post-LGCT) de cuatro ciudades mexicanas. Se realizaron análisis multivariados mediante modelos de ecuaciones de estimación generalizada (GEE). Resultados Se incremento el autoreporte de recepción de muestras gratis de cigarros (3.7% a 8.1%), ropa o artículos con marcas o logos (3.6% a 6.4%), haber visto información sobre eventos especiales (1.9% a 4.7%), y bares, antros y discos para mayores de edad (21.4% a 28%). Se observaron decrementos de publicidad en exteriores (54.7% a 47.2%). Conclusión Es necesaria una política integral con prohibiciones totales de la publicidad y promoción de los productos de tabaco que integre mayor vigilancia y sanciones para lograr la disminución y prevención del consumo de tabaco. PMID:22689158

  8. Structure of the mexicain-E-64 complex and comparison with other cysteine proteases of the papain family.

    PubMed

    Gavira, J A; González-Ramírez, L A; Oliver-Salvador, M C; Soriano-García, M; García-Ruiz, J M

    2007-05-01

    Mexicain is a 23.8 kDa cysteine protease from the tropical plant Jacaratia mexicana. It is isolated as the most abundant product after cation-exchange chromatography of the mix of proteases extracted from the latex of the fruit. The purified enzyme inhibited with E-64 [N-(3-carboxyoxirane-2-carbonyl)-leucyl-amino(4-guanido)butane] was crystallized by sitting-drop vapour diffusion and the structure was solved by molecular replacement at 2.1 A resolution and refined to an R factor of 17.7% (R(free) = 23.8%). The enzyme belongs to the alpha+beta class of proteins and the structure shows the typical papain-like fold composed of two domains, the alpha-helix-rich (L) domain and the beta-barrel-like (R) domain, separated by a groove containing the active site formed by residues Cys25 and His159, one from each domain. The four monomers in the asymmetric unit show one E-64 molecule covalently bound to Cys25 in the active site and differences have been found in the placement of E-64 in each monomer. PMID:17452780

  9. Host associations and incidence of Diuraphis spp. in the Rocky Mountain region of the United States, and pictorial key for their identification.

    PubMed

    Puterka, Gary J; Hammon, Robert W; Burd, John D; Peairs, Frank B; Randolph, Terri; Cooper, W Rodney

    2010-10-01

    The Russian wheat aphid, Diuraphis noxia Kurdjumov, is an introduced species first identified in 1986 into the United States. It has since become a major pest of wheat, Triticum aestivum L., and other small grains in the western United States. Three other Diuraphis species, Diuraphis frequens (Walker), Diuraphis mexicana (McVicar Baker), and Diuraphis tritici (Gillette), were already endemic to the United States before the introduction of D. noxia. The objective of this study was to determine the occurrence and host associations of these four Diuraphis spp. in the Rocky Mountain region that borders the western Great Plains to better understand their distribution and ecological interactions. In addition, a key to these species with photographs of live or fresh preparations of specimens is presented to aid in their identification. D. noxia was the most widely distributed species in the study area spanning the Rocky Mountain areas of Wyoming, New Mexico, Utah, and Colorado. This species was most common in the cereal-producing areas of the Colorado Plateau ecoregion. D. frequens was found to be the predominant species in the Alpine/Aspen Mountain areas of the South Central Rockies and Colorado Rockies ecoregions. The other Diuraphis species were rarely encountered even though their plant hosts occurred in the ecoregions sampled. D. noxia shared common hosts and was found co-infesting grasses with other Diuraphis species. Therefore, the potential exists for D. noxia to impact the other native Diuraphis species. PMID:21061992

  10. Extracellular polymeric substances, microbial activity and microbial community of biofilm and suspended sludge at different divalent cadmium concentrations.

    PubMed

    Wang, Zichao; Gao, Mengchun; Wei, Junfeng; Ma, Kedong; Zhang, Jing; Yang, Yusuo; Yu, Shuping

    2016-04-01

    The differences between biofilm and suspended sludge (S-sludge) in extracellular polymeric substances (EPS), microbial activity, and microbial community in an anoxic-aerobic sequencing batch biofilm reactor (SBBR) at different concentrations of divalent cadmium (Cd(II)) were investigated. As the increase of Cd(II) concentration from 0 to 50mgL(-1), the specific ammonium oxidation rate (SAOR), specific nitrite oxidation rate (SNOR), and specific nitrate reduction rate (SNRR) of biofilm decreased from 4.85, 5.22 and 45mgNg(-1) VSSh(-1) to 1.54, 2.38 and 26mgNg(-1)VSSh(-1), respectively, and the SAOR, SNOR and SNRR of S-sludge decreased from 4.80, 5.02 and 34mgNg(-1)VSSh(-1) to 1.46, 2.20 and 17mgNg(-1)VSSh(-1), respectively. Biofilm had higher protein (PN) content in EPS than S-sludge. Contrast to S-sludge, biofilm could provide Nitrobacter vulgaris, beta proteobacterium INBAF015, and Pseudoxanthomonas mexicana with the favorable conditions of growth and reproduction. PMID:26829529

  11. Recovery of perennial vegetation in military target sites in the eastern Mohave Desert, Arizona

    USGS Publications Warehouse

    Steiger, John W.; Webb, Robert H.

    2000-01-01

    The effect of the age of geomorphic surfaces on the recovery of desert vegetation in military target sites was studied in the Mohave and Cerbat Mountains of northwestern Arizona. The target sites were cleared of all vegetation during military exercises in 1942-1943 and have not been subsequently disturbed. The degree of recovery was measured by calculating percentage-similarity (PS) and correlation-coefficient indices on the basis of differences in cover, density, and volume of species growing in and out of each target site. PS values, ranging from 22.7 to 95.1 percent (100 percent = identical composition), indicate a wide range of recovery that is partially controlled by the edaphic properties of the geomorphic surfaces. Statistical analyses show a strong pattern that indicates a greater variability in the degree of recovery for sites on older surfaces than on younger surfaces and a weak pattern that indicates an inverse relation between the degree of recovery and geomorphic age. Comparisons of the different effects of target site construction on the edaphic characteristics of each target site provides an explanation for these patterns and suggests the soil properties critical to the recovery process. Statistically significant negative or positive response to disturbance for most species are independent of the age of the geomorphic surfaces; however, there is strong evidence for a shift in response for the common perennial species Acamptopappus sphaerocephalus, and to a lesser extent, Salazaria mexicana, Encelia farinosa, and Coldenia canescens, among different geomorphic surfaces.

  12. Plants used in Guatemala for the treatment of dermatophytic infections. 1. Screening for antimycotic activity of 44 plant extracts.

    PubMed

    Caceres, A; Lopez, B R; Giron, M A; Logemann, H

    1991-03-01

    Skin infections are common diseases in developing countries, of which dermatophytoses are of particular concern in the tropics, especially in infants. Through ethnobotanical surveys and literature review 100 plants were detected as being used in Guatemala for the treatment of dermatophytoses. Of these, 44 plants were screened for in vitro activity against the most common dermatophytes (Epidermophyton floccosum, Microsporum canis, Microsporum gypseum, Trichophyton mentagrophytes and Trichophyton rubrum). Results showed that aqueous extracts from 22 of the plants tested inhibit one or more of the dermatophytes. The most commonly inhibited dermatophytes were E. floccosum (43.2%), T. rubrum (36.0%), and T. mentagrophytes (31.8%); the less inhibited were M. canis (22.7%) and M. gypseum (24.0%). Plants of American origin which exhibited anti-dermatophyte activity were: Byrsonima crassifolia, Cassia grandis, Cassia occidentalis, Diphysa carthagenensis, Gliricidia sepium, Piscidia piscipula, Sambucus mexicana, Smilax regelii, Solanum americanum and Solanum nigrescens. Fungicidal and fungistatic activities as well as the minimal inhibitory concentration were demonstrated. These results provide a scientific basis for the use of these plants for the treatment of dermatophyte infections in man. PMID:2056755

  13. Trophic relationships of small nonnative fishes in a natural creek and several agricultural drains flowing into the Salton Sea, and their potential, effects on the endangered desert pupfish

    USGS Publications Warehouse

    Martin, Barbara A.; Saiki, Michael K.

    2009-01-01

    This study was conducted to characterize trophic relationships of small nonnative fishes and to determine if predation by these fishes contributes to the decline of desert pupfish (Cyprinodon macularius), an endangered cyprinodont on the verge of extinction. We sampled 403 hybrid Mozambique tilapias (Oreochromis mossambica by O. urolepis), 107 redbelly tilapias (Tilapia zillii), 32 longjaw mudsuckers (Gillkhthys mirabilis), 182 western mosquitofish (Gambusia affinis), 222 sailfin mollies (Poecilia latipinna), 63 shortfin mollies (Poecilia mexicana), and 235 porthole livebearers (Poecilurpsis gracilis) from a natural creek and four agricultural drains during September 1999- December 2001. Evidence of piscivory was in gastrointestinal contents of 14 hybrid Mozambique tilapias, 3 redbelly tilapias, 10 longjaw mudsuckers, 8 western mosquitofish, 2 sailfin mollies, and 8 porthole livebearers. Although digestion often was too advanced for identification of fishes consumed by nonnative fishes, remains of desert pupfish were in gastrointestinal contents of a longjaw mudsucker. Our findings, along with Field evidence from other studies that inverse relationships exist between abundances of desert pupfish and nonnative species, are consistent with the hypothesis that predation by nonnative species is contributing to decline of desert pupfish. We suspect that competitive interactions with nonnative fishes might also adversely affect abundance of desert pupfish.

  14. [Genetic variants associated to male infertility in Mexican patients].

    PubMed

    Piña-Aguilar, Raúl Eduardo; Chima-Galán, María del Carmen; Yerena-de-vega, María de la Concepción A; Regalado-Hernández, Miguel Angel; Sánchez-Guerrero, Cecilia; García-Ortiz, Liliana; Santillán-Hernández, Yuritzi; Moreno-García, Jesús Daniel

    2013-05-01

    Recently Mexican Federation of Obstetrics and Gynecology Colleges (Federación Mexicana de Colegios de Obstetricia y Ginecologia, FEMECOG) published the Mexican guideline forthe management of male infertility, which suggests performing genetic laboratory tests as part of diagnosis and management of infertile patients and states that these should receive genetic counseling. This paper reviews the genetic approach proposed by Mexican guideline. A systematic review of medical literature was performed in Pubmed and Web of Knowledge from 1980 to 2012 in order to find reports of genetic variants associated to male infertility in Mexican patients. Also it is discussed the current knowledge of these variants, their clinical implications and finally the guidelines and recommendations for their molecular diagnosis. Most genetic variants in Mexican infertile patients are chromosome abnormalities. In relation to other variants there is only a report of Y chromosome microdeletions, repeated CAG in androgen receptor and more common mutations in CFTR, and other article reporting mutations in CFTR in patients with congenital absence of vas deferens. Little is known about the genetics of Mexican infertile patients apart from chromosome abnormalities. However, the contribution of genetics as etiology of male infertility is taking more relevance and currently the consensual management of infertile male should include the screening of genetic background. This review pretends to be a quick guide for clinicians who want to know about reports of genetic variants related to male infertility in Mexican population and how to approach their diagnosis. PMID:23819425

  15. The impact of fallback foods on wild ring-tailed lemur biology: a comparison of intact and anthropogenically disturbed habitats.

    PubMed

    Sauther, Michelle L; Cuozzo, Frank P

    2009-12-01

    Fallback foods are often viewed as central in shaping primate morphology, and influencing adaptive shifts in hominin and other primate evolution. Here we argue that fruit of the tamarind tree (Tamarindus indica) qualifies as a fallback food of ring-tailed lemurs (Lemur catta) at the Beza Mahafaly Special Reserve (BMSR), Madagascar. Contrary to predictions that fallback foods may select for dental and masticatory morphologies adapted to processing these foods, consumption of tamarind fruit by these lemurs leaves a distinct pattern of dental pathology among ring-tailed lemurs at BMSR. Specifically, the physical and mechanical properties of tamarind fruit likely result in a high frequency of severe tooth wear, and subsequent antemortem tooth loss, in this lemur population. This pattern of dental pathology is amplified among lemurs living in disturbed areas at Beza Mahafaly, resulting from a disproportionate emphasis on challenging tamarind fruit, due to few other fruits being available. This is in part caused by a reduction in ground cover and other plants due to livestock grazing. As such, tamarind trees remain one of the few food resources in many areas. Dental pathologies are also associated with the use of a nonendemic leaf resource Argemone mexicana, an important food during the latter part of the dry season when overall food availability is reduced. Such dental pathologies at Beza Mahafaly, resulting from the use or overemphasis of fallback foods for which they are not biologically adapted, indicate that anthropogenic factors must be considered when examining fallback foods. PMID:19890872

  16. Current Knowledge of Leishmania Vectors in Mexico: How Geographic Distributions of Species Relate to Transmission Areas

    PubMed Central

    González, Camila; Rebollar-Téllez, Eduardo A.; Ibáñez-Bernal, Sergio; Becker-Fauser, Ingeborg; Martínez-Meyer, Enrique; Peterson, A. Townsend; Sánchez-Cordero, Víctor

    2011-01-01

    Leishmaniases are a group of vector-borne diseases with different clinical manifestations caused by parasites transmitted by sand fly vectors. In Mexico, the sand fly Lutzomyia olmeca olmeca is the only vector proven to transmit the parasite Leishmania mexicana to humans, which causes leishmaniasis. Other vector species with potential medical importance have been obtained, but their geographic distributions and relation to transmission areas have never been assessed. We modeled the ecological niches of nine sand fly species and projected niches to estimate potential distributions by using known occurrences, environmental coverages, and the algorithms GARP and Maxent. All vector species were distributed in areas with known recurrent transmission, except for Lu. diabolica, which appeared to be related only to areas of occasional transmission in northern Mexico. The distribution of Lu. o. olmeca does not overlap with all reported cutaneous leishmaniasis cases, suggesting that Lu. cruciata and Lu. shannoni are likely also involved as primary vectors in those areas. Our study provides useful information of potential risk areas of leishmaniasis transmission in Mexico. PMID:22049037

  17. Speciation in caves: experimental evidence that permanent darkness promotes reproductive isolation.

    PubMed

    Riesch, Rüdiger; Plath, Martin; Schlupp, Ingo

    2011-12-23

    Divergent selection through biotic factors like predation or parasitism can promote reproductive isolation even in the absence of geographical barriers. On the other hand, evidence for a role of adaptation to abiotic factors during ecological speciation in animals is scant. In particular, the role played by perpetual darkness in establishing reproductive isolation in cave animals (troglobites) remains elusive. We focused on two reproductively isolated ecotypes (surface- and cave-dwelling) of the widespread livebearer Poecilia mexicana, and raised offspring of wild-caught females to sexual maturity in a 12-month common-garden experiment. Fish were reared in light or darkness combined with high- or low-food conditions. Females, but not males, of the surface ecotype suffered from almost complete reproductive failure in darkness, especially in the low-food treatment. Furthermore, surface fish suffered from a significantly higher rate of spontaneous, stress-related infection with bacterial columnaris disease. This experimental evidence for strong selection by permanent darkness on non-adapted surface-dwelling animals adds depth to our understanding of the selective forces establishing and maintaining reproductive isolation in cave faunas. PMID:21561964

  18. Comparative in-silico genome analysis of Leishmania (Leishmania) donovani: A step towards its species specificity

    PubMed Central

    S., Satheesh Kumar; R.K., Gokulasuriyan; Ghosh, Monidipa

    2014-01-01

    Comparative genome analysis of recently sequenced Leishmania (L.) donovani was unexplored so far. The present study deals with the complete scanning of L. (L.) donovani genome revealing its interspecies variations. 60 distinctly present genes in L. (L.) donovani were identified when the whole genome was compared with Leishmania (L.) infantum. Similarly 72, 159, and 265 species specific genes were identified in L. (L.) donovani when compared to Leishmania (L.) major, Leishmania (L.) mexicana and Leishmania (Viannia) braziliensis respectively. The cross comparison of L. (L.) donovani in parallel with the other sequenced species of leishmanial led to the identification of 55 genes which are highly specific and expressed exclusively in L. (L.) donovani. We found mainly the discrepancies of surface proteins such as amastins, proteases, and peptidases. Also 415 repeat containing proteins in L. (L.) donovani and their differential distribution in other leishmanial species were identified which might have a potential role during pathogenesis. The genes identified can be evaluated as drug targets for anti-leishmanial treatment, exploring the scope for extensive future investigations. PMID:25606461

  19. Cytotoxic Effects of Bangladeshi Medicinal Plant Extracts

    PubMed Central

    Uddin, Shaikh J.; Grice, I. Darren; Tiralongo, Evelin

    2011-01-01

    To investigate the cytotoxic effect of some Bangladeshi medicinal plant extracts, 16 Bangladeshi medicinal plants were successively extracted with n-hexane, dichloromethane, methanol and water. The methanolic and aqueous extracts were screened for cytotoxic activity against healthy mouse fibroblasts (NIH3T3) and three human cancer-cell lines (gastric: AGS; colon: HT-29; and breast: MDA-MB-435S) using the MTT assay. Two methanolic extracts (Hygrophila auriculata and Hibiscus tiliaceous) and one aqueous extract (Limnophila indica) showed no toxicity against healthy mouse fibroblasts, but selective cytotoxicity against breast cancer cells (IC50 1.1–1.6 mg mL−1). Seven methanolic extracts from L. indica, Clerodendron inerme, Cynometra ramiflora, Xylocarpus moluccensis, Argemone mexicana, Ammannia baccifera and Acrostichum aureum and four aqueous extracts from Hygrophila auriculata, Bruguiera gymnorrhiza, X. moluccensis and Aegiceras corniculatum showed low toxicity (IC50 > 2.5 mg mL−1) against mouse fibroblasts but selective cytotoxicity (IC50 0.2–2.3 mg mL−1) against different cancer cell lines. The methanolic extract of Blumea lacera showed the highest cytotoxicity (IC50 0.01–0.08 mg mL−1) against all tested cell lines among all extracts tested in this study. For some of the plants their traditional use as anticancer treatments correlates with the cytotoxic results, whereas for others so far unknown cytotoxic activities were identified. PMID:19706693

  20. Shoreline foraminiferal thanatacoenoses around five eastern Caribbean islands and their environmental and biogeographic implications

    NASA Astrophysics Data System (ADS)

    Wilson, Brent; Wilson, Jacqueline I.

    2011-05-01

    Foraminiferal thanatacoenoses were examined around five islands in the Caribbean Sea, which forms a single biogeographic province with respect to nearshore (<3 m water depth) foraminifera, which live primarily on marine vegetation. On death, they become incorporated into the sediment. The assemblage at a site reflects the live assemblage and post mortem affects such as dissolution and transport during storms. Reefal species ( Amphistegina gibbosa, Asterigerina carinata) are transported towards shore during storms. Foraminiferal thanatacoenoses were examined in 65 nearshore sediment samples from around five eastern Caribbean islands: St. John (US Virgin Islands), St. Kitts, Nevis, Bequia and Tobago. Cluster and principal components analyses distinguished the following environments (indicator species in parentheses): Sediment associated with mangroves ( Ammonia sobrina), Bays subject to organic matter enrichment ( Quinqueloculina poeyana, Triloculina rotunda, T. trigonula), Areas subject to moderate sediment flux during storms ( Quinqueloculina auberiana, Nodobaculariella mexicana, Peneroplis proteus, Archaias angulatus), Locations subject to high sediment flux during storms ( Amphistegina gibbosa), Sites little stressed by organic matter enrichment or storms ( Discorbis rosea). The majority of samples were from sites in the last category. The data from this study could form the nucleus of a catalogue of Caribbean beaches and their environmental influences.

  1. Single-molecule analysis of DNA replication reveals novel features in the divergent eukaryotes Leishmania and Trypanosoma brucei versus mammalian cells

    PubMed Central

    Stanojcic, Slavica; Sollelis, Lauriane; Kuk, Nada; Crobu, Lucien; Balard, Yves; Schwob, Etienne; Bastien, Patrick; Pagès, Michel; Sterkers, Yvon

    2016-01-01

    Leishmania and Trypanosoma are unicellular parasites that possess markedly original biological features as compared to other eukaryotes. The Leishmania genome displays a constitutive ‘mosaic aneuploidy’, whereas in Trypanosoma brucei, the megabase-sized chromosomes are diploid. We accurately analysed DNA replication parameters in three Leishmania species and Trypanosoma brucei as well as mouse embryonic fibroblasts (MEF). Active replication origins were visualized at the single molecule level using DNA molecular combing. More than one active origin was found on most DNA fibres, showing that the chromosomes are replicated from multiple origins. Inter-origin distances (IODs) were measured and found very large in trypanosomatids: the mean IOD was 160 kb in T. brucei and 226 kb in L. mexicana. Moreover, the progression of replication forks was faster than in any other eukaryote analyzed so far (mean velocity 1.9 kb/min in T. brucei and 2.4–2.6 kb/min in Leishmania). The estimated total number of active DNA replication origins in trypanosomatids is ~170. Finally, 14.4% of unidirectional replication forks were observed in T. brucei, in contrast to 1.5–1.7% in Leishmania and 4% in MEF cells. The biological significance of these original features is discussed. PMID:26976742

  2. Effect of psilocin on extracellular dopamine and serotonin levels in the mesoaccumbens and mesocortical pathway in awake rats.

    PubMed

    Sakashita, Yuichi; Abe, Kenji; Katagiri, Nobuyuki; Kambe, Toshie; Saitoh, Toshiaki; Utsunomiya, Iku; Horiguchi, Yoshie; Taguchi, Kyoji

    2015-01-01

    Psilocin (3-[2-(dimethylamino)ethyl]-1H-indol-4-ol) is a hallucinogenic component of the Mexican mushroom Psilocybe mexicana and a skeletal serotonin (5-HT) analogue. Psilocin is the active metabolite of psilocybin (3-[2-(dimethylamino)ethyl]-1H-indol-4-yl dihydrogen phosphate). In the present study, we examined the effects of systemically administered psilocin on extracellular dopamine and 5-HT concentrations in the ventral tegmental area (VTA), nucleus accumbens, and medial prefrontal cortex of the dopaminergic pathway in awake rats using in vivo microdialysis. Intraperitoneal administration of psilocin (5, 10 mg/kg) significantly increased extracellular dopamine levels in the nucleus accumbens. Psilocin did not affect the extracellular 5-HT level in the nucleus accumbens. Conversely, systemic administration of psilocin (10 mg/kg) significantly increased extracellular 5-HT levels in the medial prefrontal cortex of rats, but dopamine was decreased in this region. However, neither extracellular dopamine nor 5-HT levels in the VTA were altered by administration of psilocin. Behaviorally, psilocin significantly increased the number of head twitches. Thus, psilocin affects the dopaminergic system in the nucleus accumbens. In the serotonergic system, psilocin contribute to a crucial effect in the medial prefrontal cortex. The present data suggest that psilocin increased both the extracellular dopamine and 5-HT concentrations in the mesoaccumbens and/or mesocortical pathway. PMID:25342005

  3. Atlas of Mexican Triatominae (Reduviidae: Hemiptera) and vector transmission of Chagas disease

    PubMed Central

    Ramsey, Janine M; Peterson, A Townsend; Carmona-Castro, Oscar; Moo-Llanes, David A; Nakazawa, Yoshinori; Butrick, Morgan; Tun-Ku, Ezequiel; de la Cruz-Félix, Keynes; Ibarra-Cerdeña, Carlos N

    2015-01-01

    Chagas disease is one of the most important yet neglected parasitic diseases in Mexico and is transmitted by Triatominae. Nineteen of the 31 Mexican triatomine species have been consistently found to invade human houses and all have been found to be naturally infected with Trypanosoma cruzi. The present paper aims to produce a state-of-knowledge atlas of Mexican triatomines and analyse their geographic associations with T. cruzi, human demographics and landscape modification. Ecological niche models (ENMs) were constructed for the 19 species with more than 10 records in North America, as well as for T. cruzi. The 2010 Mexican national census and the 2007 National Forestry Inventory were used to analyse overlap patterns with ENMs. Niche breadth was greatest in species from the semiarid Nearctic Region, whereas species richness was associated with topographic heterogeneity in the Neotropical Region, particularly along the Pacific Coast. Three species, Triatoma longipennis, Triatoma mexicana and Triatoma barberi, overlapped with the greatest numbers of human communities, but these communities had the lowest rural/urban population ratios. Triatomine vectors have urbanised in most regions, demonstrating a high tolerance to human-modified habitats and broadened historical ranges, exposing more than 88% of the Mexican population and leaving few areas in Mexico without the potential for T. cruzi transmission. PMID:25993505

  4. Systematics of disakisperma (poaceae, chloridoideae, chlorideae).

    PubMed

    Snow, Neil; Peterson, Paul M; Romaschenko, Konstantin

    2013-01-01

    Disakisperma Steud. is a genus of four predominantly perennial C4 (NAD-ME) species in the Americas, Africa, and Asia. Its species previously were treated in Eleusine, Eragrostis, Coelachyrum, Cypholepis, Leptochloa, or Diplachne by nearly all authors.It includes the widespread North and South American amphitropical disjunct Disakisperma dubium (type of the genus), Disakisperma eleusine from southern Africa, Disakisperma obtusiflorum from central and northern Africa to southern Asia, and Disakisperma yemenicum, comb. nov. from eastern and southern Africa to Yemen. This paper provides a key to the species, geographic distributions, descriptions, including comments on the anatomy of leaves, stems, lemmatal micromorphology, a phylogram based on five molecular markers, and discussions of chromosome numbers. The species are rarely, if at all, known outside of their native ranges and are unlikely to become aggressively invasive. All species are considered Least Concern following IUCN guidelines. Lectotypes are designated for Diplachne dubia var. pringleana Kuntze, Disakisperma mexicana Steud., Eragrostis yemenica Schweinf., and Leptochloa appletonii Stapf. PMID:24194669

  5. Reduction of Energetic Demands through Modification of Body Size and Routine Metabolic Rates in Extremophile Fish.

    PubMed

    Passow, Courtney N; Greenway, Ryan; Arias-Rodriguez, Lenin; Jeyasingh, Punidan D; Tobler, Michael

    2015-01-01

    Variation in energy availability or maintenance costs in extreme environments can exert selection for efficient energy use, and reductions in organismal energy demand can be achieved in two ways: reducing body mass or metabolic suppression. Whether long-term exposure to extreme environmental conditions drives adaptive shifts in body mass or metabolic rates remains an open question. We studied body size variation and variation in routine metabolic rates in locally adapted populations of extremophile fish (Poecilia mexicana) living in toxic, hydrogen sulfide-rich springs and caves. We quantified size distributions and routine metabolic rates in wild-caught individuals from four habitat types. Compared with ancestral populations in nonsulfidic surface habitats, extremophile populations were characterized by significant reductions in body size. Despite elevated metabolic rates in cave fish, the body size reduction precipitated in significantly reduced energy demands in all extremophile populations. Laboratory experiments on common garden-raised fish indicated that elevated routine metabolic rates in cave fish likely have a genetic basis. The results of this study indicate that adaptation to extreme environments directly impacts energy metabolism, with fish living in cave and sulfide spring environments expending less energy overall during routine metabolism. PMID:26052634

  6. Phylogenetic overview of the genus Genea (Pezizales, Ascomycota) with an emphasis on European taxa.

    PubMed

    Alvarado, Pablo; Cabero, Julio; Moreno, Gabriel; Bratek, Zoltán; van Vooren, Nicolas; Kaounas, Vasileios; Konstantinidis, Giorgos; Agnello, Carlo; Merényi, Zsolt; Smith, Matthew E; Vizzini, Alfredo; Trappe, James M

    2016-01-01

    We constructed a comprehensive phylogeny of the genus Genea, with new molecular data from samples collected in several countries in temperate and Mediterranean Europe, as well as North America. Type specimens and authentic material of most species were examined to support identifications. The molecular identity of the most common species in Genea was compared with nuc rDNA internal transcribed spacer (ITS), D1-D2 domains of 28S nuc rDNA (28S rDNA) and translation elongation factor 1-α ene (TEF1) profiles of 10 recently proposed taxa, G. brunneocarpa, G. compressa, G. dentata, G. fageticola, G. lobulata, G. oxygala, G. pinicola, G. pseudobalsleyi, G. pseudoverrucosa and G. tuberculata, supporting their status as distinct species. Genea mexicana and G. thaxteri on the one hand and G. sphaerica and G. lespiaultii on the other are closely related. Multiple lineages were recorded for G. verrucosa and G. fragrans, but we found no morphological traits to discriminate among them, so we tentatively interpreted them as cryptic species. A key to species of the genus Genea is provided to facilitate identification. We provide macroscopic images of fresh specimens and of representative spores of most species. Finally, we conducted a molecular analysis of the divergence time for Genea and discuss the implications of our results. PMID:26740538

  7. Optimizing conservation strategies for Mexican freetailed bats: a population viability and ecosystem services approach

    USGS Publications Warehouse

    Wiederholt, Ruscena; Lopez-Hoffman, Laura; Svancara, Colleen; McCracken, Gary; Thogmartin, Wayne E.; Diffendorfer, James E.; Mattson, Brady; Bagstad, Kenneth J.; Cryan, Paul; Amy Russell; Semmens, Darius J.; Rodrigo A. Medellín

    2015-01-01

    Conservation planning can be challenging due to the need to balance biological concerns about population viability with social concerns about the benefits biodiversity provide to society, often while operating under a limited budget. Methods and tools that help prioritize conservation actions are critical for the management of at-risk species. Here, we use a multi-attribute utility function to assess the optimal maternity roosts to conserve for maintaining the population viability and the ecosystem services of a single species, the Mexican free-tailed bat (Tadarida brasiliensis mexicana). Mexican free-tailed bats provide ecosystem services such as insect pest-suppression in agricultural areas and recreational viewing opportunities, and may be threatened by climate change and development of wind energy. We evaluated each roost based on five attributes: the maternity roost’s contribution to population viability, the pest suppression ecosystem services to the surrounding area provided by the bats residing in the roost, the ecotourism value of the roost, the risks posed to each roost structure, and the risks posed to the population of bats residing in each roost. We compared several scenarios that prioritized these attributes differently, hypothesizing that the set of roosts with the highest rankings would vary according to the conservation scenario. Our results indicate that placing higher values on different roost attributes (e.g. population importance over ecosystem service value) altered the roost rankings. We determined that the values placed on various conservation objectives are an important determinant of habitat planning.

  8. Taxonomy, diversity, temporal and geographical distribution of Cutaneous Leishmaniasis in Colombia: A retrospective study.

    PubMed

    Ramírez, Juan David; Hernández, Carolina; León, Cielo M; Ayala, Martha S; Flórez, Carolina; González, Camila

    2016-01-01

    Leishmaniases are tropical zoonotic diseases, caused by kinetoplastid parasites from the genus Leishmania. New World (NW) species are related to sylvatic cycles although urbanization processes have been reported in some South American Countries such as Colombia. Currently, few studies show the relative distribution of Leishmania species related to cutaneous Leishmaniasis (CL) in South America due to the lack of accurate surveillance and public health systems. Herein, we conducted a systematic estimation of the Leishmania species causing CL in Colombia from 1980 to 2001 via molecular typing and isoenzymes. A total of 327 Leishmania isolates from humans, sandflies and reservoirs were typed as L. panamensis 61.3% (201), L. braziliensis 27.1% (88), L. lainsoni 0.6% (2), L. guyanensis 0.9% (3), L. infantum chagasi 4% (12), L. equatoriensis 0.6% (2), L. mexicana 2.1% (8), L. amazonensis 2.8% (9) and L. colombiensis 0.6% (2). This is the first report of two new Leishmania species circulating in Colombia and suggests the need to convince the Colombian government about the need to deploy and standardize tools for the species identification to provide adequate management to individuals suffering this pathology. PMID:27328969

  9. Effect of biotic factors on the spatial distribution of stingless bees (Hymenoptera: Apidae, Meliponini) in fragmented neotropical habitats.

    PubMed

    Fierro, M M; Cruz-López, L; Sánchez, D; Villanueva-Gutiérrez, R; Vandame, R