text:
Contact Us
Enter Your Search:
Search tips
Search criteria
Journal titles
Results 1-25 (311775)
« Previous
Go to page number:
Next »
Clipboard (0)
Related Articles
Overexpression of the Natural Inhibitor of Cysteine Peptidases in Leishmania mexicana Leads to Reduced Virulence and a Th1 Response▿
Bryson, Karen
Besteiro, Sébastien
McGachy, H. Adrienne
Coombs, Graham H.
Mottram, Jeremy C.
Alexander, James
Infection and Immunity
Leishmania mexicana cysteine peptidases (CPs) have been identified as important parasite virulence factors. More recently, a natural inhibitor of CPs (ICP) from L. mexicana has been characterized, and ICP mutants have been created. Infection of BALB/c mice with ICP null mutants or ICP reexpressing mutants resulted in nonhealing, progressively growing lesions albeit slightly attenuated compared with the growth of lesions produced by wild-type parasites. In contrast, BALB/c mice infected with mutants overexpressing ICP were able to significantly control lesion growth or heal. While BALB/c mice infected with wild-type parasites, ICP null mutants, or ICP reexpressing mutants produced significant antibody responses, including immunoglobulin E (IgE), no Th1 response, as indicated by antigen-induced splenocyte gamma interferon (IFN-γ) production, could be demonstrated. In contrast, BALB/c mice infected with mutants overexpressing ICP produced significantly less antibody, particularly IgE, as well as significantly reduced splenocyte interleukin-4 and enhanced IFN-γ production. BALB/c mice were able to resolve infection following infection with one ICP overexpressing clone, which was subsequently used for vaccination studies with BALB/c mice. However, no protection was afforded these mice when they were challenged with wild-type parasites. Nevertheless, two other mouse strains susceptible to L. mexicana, C3H and C57BL/6, vaccinated with overexpressing ICP mutants were able to control challenge infection associated with an enhanced Th1 response. This study confirms that L. mexicana CPs are virulence factors and that ICPs have therapeutic potential.
Related Articles
A potential role for ICP, a leishmanial inhibitor of cysteine peptidases, in the interaction between host and parasite
Besteiro, Sébastien
Coombs, Graham H.
Mottram, Jeremy C.
Molecular microbiology
The biological role of a natural inhibitor of cysteine peptidases (designated ICP) of Leishmania has been investigated by genetic manipulation of the parasite. Null mutants grew normally in vitro, were as infective to macrophages in vitro as wild-type parasites, but had reduced infectivity to mice. Mutants re-expressing ICP from a single gene gave partial restoration of virulence in vivo, whereas mutants over-expressing ICP secreted the inhibitor and showed markedly reduced virulence in mice. Promastigotes of the null mutants had similar cysteine peptidase activities as the wild-type parasites, suggesting that ICP is not required for the expression or processing of the enzymes. The only proteins found to bind to ICP in promastigote cell lysates were fully processed forms of CPA and CPB, showing that ICP does not bind in abundance either to zymogens of the cysteine peptidases or other leishmanial proteins. However, only a small proportion of ICP co-localised with CPA and CPB in the promastigote (in the endoplasmic reticulum and Golgi) and the majority of ICP resided in vesicles that are apparently distinct from endosomes and the multivesicular tubule (MVT)-lysosome. These data suggest that ICP has a role other than modulation of the activity of the parasite's own cysteine peptidases and their normal trafficking to the MVT-lysosome via the flagellar pocket. The finding that ICP partially co-localised with an endocytosed cysteine peptidase leads us to postulate that ICP has a role in protection of the parasite against the hydrolytic environment of the sandfly gut and/or the parasitophorous vacuole of host macrophages.
cysteine peptidase inhibitor; ICP; chagasin; lysosomes; endosomes; Leishmania
Related Articles
Smith, Brian O.
Picken, Nichola C.
Westrop, Gareth D.
Bromek, Krystyna
Mottram, Jeremy C.
Coombs, Graham H.
The Journal of biological chemistry
Clan CA, family C1 cysteine peptidases (CPs) are important virulence factors and drug targets in parasites that cause neglected diseases. Natural CP inhibitors of the I42 family, known as ICP, occur in some protozoa and bacterial pathogens, but are absent from metazoa. They are active against both parasite and mammalian CPs, despite having no sequence similarity with other classes of CP inhibitor. Recent data suggest that L. mexicana ICP plays an important role in host-parasite interactions. We have now solved the structure of ICP from L. mexicanaby NMR and shown that it adopts a type of immunoglobulin-like fold not previously reported in lower eukaryotes or bacteria. The structure places three loops containing highly conserved residues at one end of the molecule, one loop being highly mobile. Interaction studies with CPs confirm the importance of these loops for the interaction between ICP and CPs and suggest the mechanism of inhibition. Structure-guided mutagenesis of ICP has revealed that residues in the mobile loop are critical for CP inhibition. Data-driven docking models support the importance of the loops in the ICP-CP interaction. This study provides structural evidence for the convergent evolution from an immunoglobulin fold of CP inhibitors with a cystatin-like mechanism.
Related Articles
Cathepsin-L Can Resist Lysis by Human Serum in Trypanosoma brucei brucei
Alsford, Sam
Currier, Rachel B.
Guerra-Assunção, José Afonso
Clark, Taane G.
Horn, David
PLoS Pathogens
Closely related African trypanosomes cause lethal diseases but display distinct host ranges. Specifically, Trypanosoma brucei brucei causes nagana in livestock but fails to infect humans, while Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense cause sleeping sickness in humans. T. b. brucei fails to infect humans because it is sensitive to innate immune complexes found in normal human serum known as trypanolytic factor (TLF) 1 and 2; the lytic component is apolipoprotein-L1 in both TLFs. TLF resistance mechanisms of T. b. gambiense and T. b. rhodesiense are now known to arise through either gain or loss-of-function, but our understanding of factors that render T. b. brucei susceptible to lysis by human serum remains incomplete. We conducted a genome-scale RNA interference (RNAi) library screen for reduced sensitivity to human serum. Among only four high-confidence 'hits' were all three genes previously shown to sensitize T. b. brucei to human serum, the haptoglobin-haemoglobin receptor (HpHbR), inhibitor of cysteine peptidase (ICP) and the lysosomal protein, p67, thereby demonstrating the pivotal roles these factors play. The fourth gene identified encodes a predicted protein with eleven trans-membrane domains. Using chemical and genetic approaches, we show that ICP sensitizes T. b. brucei to human serum by modulating the essential cathepsin, CATL, a lysosomal cysteine peptidase. A second cathepsin, CATB, likely to be dispensable for growth in in vitro culture, has little or no impact on human-serum sensitivity. Our findings reveal major and novel determinants of human-serum sensitivity in T. b. brucei. They also shed light on the lysosomal protein-protein interactions that render T. b. brucei exquisitely sensitive to lytic factors in human serum, and indicate that CATL, an important potential drug target, has the capacity to resist these factors.
Author Summary
The interplay among host innate immunity and resistance mechanisms in African trypanosomes has a major impact on the host range of these tsetse-fly transmitted parasites, defining their ability to cause disease in humans. A genome-scale RNAi screen identified a highly restricted set of four genes that sensitise trypanosomes to human serum: those encoding the haptoglobin-haemoglobin receptor, a predicted trans-membrane channel, a lysosomal membrane-protein and the cysteine peptidase inhibitor. An analysis of the cysteine peptidases revealed cathepsin-L as the protease regulated by the inhibitor – and with the capacity to render the parasite resistant to lysis by human serum. These findings emphasise the importance of parasite factors for the delivery and stability of host toxins. They also shed light on the control of proteolysis by parasites and potential unanticipated consequences of therapies that target the parasite proteases.
Related Articles
Leishmania mexicana promastigotes inhibit macrophage IL-12 production via TLR-4 dependent COX-2, iNOS and arginase-1 expression
Shweash, Muhannad
Adrienne McGachy, H.
Schroeder, Juliane
Neamatallah, Thikryat
Bryant, Clare E.
Millington, Owain
Mottram, Jeremy C.
Alexander, James
Plevin, Robin
Molecular Immunology
► TLR-4 activation by Leishmania mexicana promastigotes and CPB-deficient amastigotes. ► Prolonged, TLR-4 dependent iNOS and COX-2 expression by L. mexicana promastigotes. ► Enhanced TLR-4 dependent-arginase-1 expression. ► Regulation of IL-12 induction by an arginase-1 dependent mechanism.
The effects of Leishmania mexicana metacyclic promastigotes upon MAP kinase signalling in mouse bone marrow macrophages and subsequent expression of the disease regulatory proteins iNOS and COX-2 were studied. At a ratio of 5:1, promastigotes caused a marked increase in phosphorylation of the three major MAP kinases, ERK, p38 and JNK. MAP kinase signalling was substantially reduced in TLR-4−/− but not TLR-2−/− deficient macrophages and completely abolished in double TLR-2/4−/− macrophages. A similar outcome was observed using cysteine peptidase B deficient amastigotes. Furthermore, whilst promastigotes had no independent effect on iNOS or COX-2 expression, they prolonged the induction of these proteins stimulated by LPS and enhanced PGE2 and NO production. Induction of COX-2 and iNOS was also TLR-4 dependent. Blockade of either PGE2 or NO production with indomethacin or l-NAME reversed promastigote inhibition of LPS induced IL-12 production. Promastigotes also increased macrophage arginase-1 expression and enhanced arginase activity, both of which were substantially reduced in TLR-4 but not TLR-2 deficient macrophages. Surprisingly, arginase inhibition by Nor-NOHA also caused a reversal of promastigote mediated inhibition of macrophage IL-12 production. These data demonstrate for the first time the role of TLR-4 in mediating the effects of L. mexicana promastigotes on MAP kinase activation, up-regulation of COX-2, iNOS as well as arginase-1 expression in macrophages and further shows that PGE2, NO and arginase activity all contribute substantially to the inhibition of host cell IL-12 production.
Leishmania mexicana; Promastigotes; TLR-4; MAP kinase; Arginase-1; IL-12
Related Articles
Identification of Semicarbazones, Thiosemicarbazones and Triazine Nitriles as Inhibitors of Leishmania mexicana Cysteine Protease CPB
Schröder, Jörg
Noack, Sandra
Marhöfer, Richard J.
Mottram, Jeremy C.
Coombs, Graham H.
Selzer, Paul M.
PLoS ONE
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.
Related Articles
Surface Zn-proteinase as a molecule for defense of Leishmania mexicana amazonensis promastigotes against cytolysis inside macrophage phagolysosomes.
Seay, M B
Heard, P L
Chaudhuri, G
Infection and Immunity
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.
Related Articles
HIV Aspartyl Peptidase Inhibitors Interfere with Cellular Proliferation, Ultrastructure and Macrophage Infection of Leishmania amazonensis
Santos, Lívia O.
Marinho, Fernanda A.
Altoé, Ellen F.
Vitório, Bianca S.
Alves, Carlos R.
Britto, Constança
Motta, Maria Cristina M.
Branquinha, Marta H.
Santos, André L. S.
d'Avila-Levy, Claudia M.
PLoS ONE
Leishmania is the etiologic agent of leishmanisais, a protozoan disease whose pathogenic events are not well understood. Current therapy is suboptimal due to toxicity of the available therapeutic agents and the emergence of drug resistance. Compounding these problems is the increase in the number of cases of Leishmania-HIV coinfection, due to the overlap between the AIDS epidemic and leishmaniasis.
Methodology/Principal Findings
In the present report, we have investigated the effect of HIV aspartyl peptidase inhibitors (PIs) on the Leishmania amazonensis proliferation, ultrastructure, interaction with macrophage cells and expression of classical peptidases which are directly involved in the Leishmania pathogenesis. All the HIV PIs impaired parasite growth in a dose-dependent fashion, especially nelfinavir and lopinavir. HIV PIs treatment caused profound changes in the leishmania ultrastructure as shown by transmission electron microscopy, including cytoplasm shrinking, increase in the number of lipid inclusions and some cells presenting the nucleus closely wrapped by endoplasmic reticulum resembling an autophagic process, as well as chromatin condensation which is suggestive of apoptotic death. The hydrolysis of HIV peptidase substrate by L. amazonensis extract was inhibited by pepstatin and HIV PIs, suggesting that an aspartyl peptidase may be the intracellular target of the inhibitors. The treatment with HIV PIs of either the promastigote forms preceding the interaction with macrophage cells or the amastigote forms inside macrophages drastically reduced the association indexes. Despite all these beneficial effects, the HIV PIs induced an increase in the expression of cysteine peptidase b (cpb) and the metallopeptidase gp63, two well-known virulence factors expressed by Leishmania spp.
In the face of leishmaniasis/HIV overlap, it is critical to further comprehend the sophisticated interplays among Leishmania, HIV and macrophages. In addition, there are many unresolved questions related to the management of Leishmania-HIV-coinfected patients. For instance, the efficacy of therapy aimed at controlling each pathogen in coinfected individuals remains largely undefined. The results presented herein add new in vitro insight into the wide spectrum efficacy of HIV PIs and suggest that additional studies about the synergistic effects of classical antileishmanial compounds and HIV PIs in macrophages coinfected with Leishmania and HIV-1 should be performed.
Related Articles
Leishmania mexicana metacaspase is a negative regulator of amastigote proliferation in mammalian cells
Castanys-Muñoz, E
Brown, E
Coombs, G H
Mottram, J C
Cell Death & Disease
Metacaspases (MCAs) are caspase family cysteine peptidases that have been implicated in cell death processes in plants, fungi and protozoa. MCAs have also been suggested to be involved in cell cycle control, differentiation and clearance of aggregates; they are virulence factors. Dissecting the function of MCAs has been complicated by the presence in many organisms of multiple MCA genes or limitations on genetic manipulation. We describe here the creation of a MCA gene-deletion mutant (Δmca) in the protozoan parasite Leishmania mexicana, which has allowed us to dissect the role of the parasite's single MCA gene in cell growth and cell death. Δmca parasites are viable as promastigotes, and differentiate normally to the amastigote form both in in vitro macrophages infection and in mice. Δmca promastigotes respond to cell death inducers such as the drug miltefosine and H2O2 similarly to wild-type (WT) promastigotes, suggesting that MCAs do not have a caspase-like role in execution of L. mexicana cell death. Δmca amastigotes replicated significantly faster than WT amastigotes in macrophages and in mice, but not as axenic culture in vitro. We propose that the Leishmania MCA acts as a negative regulator of amastigote proliferation, thereby acting to balance cell growth and cell death.
metacaspase; caspase; cysteine peptidase; programmed cell death
Related Articles
Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5
Denise, Hubert
Poot, Jacqueline
Jiménez, Maribel
Ambit, Audrey
Herrmann, Daland C
Vermeulen, Arno N
Coombs, Graham H
Mottram, Jeremy C
BMC Molecular Biology
Visceral leishmaniasis caused by members of the Leishmania donovani complex is often fatal in the absence of treatment. Research has been hampered by the lack of good laboratory models and tools for genetic manipulation. In this study, we have characterised a L. infantum line (JPCM5) that was isolated from a naturally infected dog and then cloned. We found that JPCM5 has attributes that make it an excellent laboratory model; different stages of the parasite life cycle can be studied in vitro, it is accessible to genetic manipulation and it has retained its virulence. Furthermore, the L. infantum JPCM5 genome has now been fully sequenced.
We have further focused our studies on LiCPA, the L. infantum homologue to L. mexicana cysteine peptidase CPA. LiCPA was found to share a high percentage of amino acid identity with CPA proteins of other Leishmania species. Two independent LiCPA-deficient promastigote clones (ΔLicpa) were generated and their phenotype characterised. In contrast to L. mexicana CPA-deficient mutants, both clones of ΔLicpa were found to have significantly reduced virulence in vitro and in vivo. Re-expression of just one LiCPA allele (giving ΔLicpa::CPA) was sufficient to complement the reduced infectivity of both ΔLicpa mutants for human macrophages, which confirms the importance of LiCPA for L. infantum virulence. In contrast, in vivo experiments did not show any virulence recovery of the re-expressor clone ΔLicpaC1::CPA compared with the CPA-deficient mutant ΔLicpaC1.
The data suggest that CPA is not essential for replication of L. infantum promastigotes, but is important for the host-parasite interaction. Further studies will be necessary to elucidate the precise roles that LiCPA plays and why the re-expression of LiCPA in the ΔLicpa mutants complemented the gene deletion phenotype only in in vitro and not in in vivo infection of hamsters.
Related Articles
Comparative in-silico genome analysis of Leishmania (Leishmania) donovani: A step towards its species specificity
S., Satheesh Kumar
R.K., Gokulasuriyan
Ghosh, Monidipa
Meta Gene
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.
•Comparative genome analysis identifies 55 species specific L. (L.) donovani genes.•Discrepancies of surface proteins such as amastins, proteases, and peptidases are identified in L. (L.) donovani.•Apical Membrane Antigen (AMA1) might be a novel factor which helps L. (L.) donovani invasion.•Novel A2 and amastin genes in L. (L.) donovani genome are identified.•Our study identifies differential gene distribution in L. (L.) donovani with respect to other leishmanial species.
Visceral leishmaniasis; Apical Membrane Antigen 1; A2 gene family; Amastin; Species specific genes
Related Articles
Inhibitor of Cysteine Proteases Is Critical for Motility and Infectivity of Plasmodium Sporozoites
Boysen, Katja E.
Matuschewski, Kai
Malaria is transmitted when motile sporozoites are injected into the dermis by an infected female Anopheles mosquito. Inside the mosquito vector, sporozoites egress from midgut-associated oocysts and eventually penetrate the acinar cells of salivary glands. Parasite-encoded factors with exclusive vital roles in the insect vector can be studied by classical reverse genetics. Here, we characterized the in vivo roles of Plasmodium berghei falstatin/ICP (inhibitor of cysteine proteases). This protein was previously suggested to act as a protease inhibitor during erythrocyte invasion. We show by targeted gene disruption that loss of ICP function does not affect growth inside the mammalian host but causes a complete defect in sporozoite transmission. Sporogony occurred normally in icp(−) parasites, but hemocoel sporozoites showed a defect in continuous gliding motility and infectivity for salivary glands, which are prerequisites for sporozoite transmission to the mammalian host. Absence of ICP correlates with enhanced cleavage of circumsporozoite protein, in agreement with a role as a protease regulator. We conclude that ICP is essential for only the final stages of sporozoite maturation inside the mosquito vector. This study is the first genetic evidence that an ICP is necessary for the productive motility of a eukaryotic parasitic cell.
Cysteine proteases and their inhibitors are considered ideal drug targets for the treatment of a wide range of diseases, including cancer and parasitic infections. In protozoan parasites, including Leishmania, Trypanosoma, and Plasmodium, cysteine proteases play important roles in life cycle progression. A mouse malaria model provides an unprecedented opportunity to study the roles of a parasite-encoded inhibitor of cysteine proteases (ICP) over the entire parasite life cycle. By precise gene deletion, we found no evidence that ICP influences disease progression or parasite virulence. Instead, we discovered that this factor is necessary for parasite movement and malaria transmission from mosquitoes to mammals. This finding in a fast-moving unicellular protozoan has important implications for malaria intervention strategies and the roles of ICPs in the regulation of eukaryotic cell migration.
Related Articles
Disease Severity in Patients Infected with Leishmania mexicana Relates to IL-1β
Fernández-Figueroa, Edith A.
Rangel-Escareño, Claudia
Espinosa-Mateos, Valeria
Carrillo-Sánchez, Karol
Salaiza-Suazo, Norma
Carrada-Figueroa, Georgina
March-Mifsut, Santiago
Becker, Ingeborg
PLoS Neglected Tropical Diseases
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.
Author Summary
Leishmania mexicana is an intracellular parasite that causes two polarly opposed diseases: One is a self-limited disease, characterized by ulcerative lesions associated with a low infectious load, as found in patients with localized cutaneous leishmaniasis (LCL). And the other pole is characterized by a progressive disease where abundant parasites spread uncontrollably throughout the skin inside heavily infected phagocytic cells, as occurs in patients with diffuse cutaneous leishmaniasis (DCL). The cause of this severe form of the disease is unknown, although the early encounter between the parasite and the inflammatory response of the host possibly plays a decisive role in the disease outcome. We here show that polymorphism in the gene encoding IL-1β (−511 C/T) represents a variable influencing the risk to develop the disease for patients infected with Leishmania mexicana. In vitro experiments showed that monocytes of DCL patients secreted significantly higher levels of the proinflammatory cytokine IL-1β as compared to LCL patients. DCL patients also had augmented levels of IL-1β in serum, and the cytokine was diffusely distributed throughout lesions, which was correlated with the numbers of parasites in the lesions. We propose that IL-1β possibly plays a key role in establishing the disease severity in patients infected with Leishmania mexicana.
Related Articles
MAP Kinase Phosphatase-2 Plays a Critical Role in Response to Infection by Leishmania mexicana
Al-Mutairi, Mashael S.
Cadalbert, Laurence C.
McGachy, H. Adrienne
Shweash, Muhannad
Schroeder, Juliane
Kurnik, Magdalena
Sloss, Callum M.
Bryant, Clare E.
Alexander, James
Plevin, Robin
PLoS Pathogens
In this study we generated a novel dual specific phosphatase 4 (DUSP4) deletion mouse using a targeted deletion strategy in order to examine the role of MAP kinase phosphatase-2 (MKP-2) in immune responses. Lipopolysaccharide (LPS) induced a rapid, time and concentration-dependent increase in MKP-2 protein expression in bone marrow-derived macrophages from MKP-2+/+ but not from MKP-2−/− mice. LPS-induced JNK and p38 MAP kinase phosphorylation was significantly increased and prolonged in MKP-2−/− macrophages whilst ERK phosphorylation was unaffected. MKP-2 deletion also potentiated LPS-stimulated induction of the inflammatory cytokines, IL-6, IL-12p40, TNF-α, and also COX-2 derived PGE2 production. However surprisingly, in MKP-2−/− macrophages, there was a marked reduction in LPS or IFNγ-induced iNOS and nitric oxide release and enhanced basal expression of arginase-1, suggesting that MKP-2 may have an additional regulatory function significant in pathogen-mediated immunity. Indeed, following infection with the intracellular parasite Leishmania mexicana, MKP-2−/− mice displayed increased lesion size and parasite burden, and a significantly modified Th1/Th2 bias compared with wild-type counterparts. However, there was no intrinsic defect in MKP-2−/− T cell function as measured by anti-CD3 induced IFN-γ production. Rather, MKP-2−/− bone marrow-derived macrophages were found to be inherently more susceptible to infection with Leishmania mexicana, an effect reversed following treatment with the arginase inhibitor nor-NOHA. These findings show for the first time a role for MKP-2 in vivo and demonstrate that MKP-2 may be essential in orchestrating protection against intracellular infection at the level of the macrophage.
Author Summary
In cells of the immune system are switch-on enzymes called kinases which regulate responses to infectious agents such as Leishmania. However, in the same cells there are switch-off enzymes known as phosphatases which function to turn off the kinases once they have done their work. A lot of studies have focussed on the role of kinases but not phosphatases in response to infection; we therefore generated a novel mouse in which the gene for one of these phosphatases, called MKP-2, has been deleted. We found that in the absence of this phosphatase unexpected things happened. The profile of inflammatory proteins, produced by a special cell of the immune system, called a macrophage, that functions to regulate infection by Leishmania, changed in ways which meant the macrophage could either fight infection very effectively or very weakly. In actual fact, we found that the macrophages with no MKP-2 fought off Leishmania poorly and mice deficient in MKP-2 had a modified immune response favouring the growth of the parasite. This is the first study to give critical insight into the role of MKP-2 in fighting Leishmania infection and demonstrates very well the importance of this class of enzyme in pathogen biology.
Related Articles
Nelfinavir is effective in inhibiting the multiplication and aspartic peptidase activity of Leishmania species, including strains obtained from HIV-positive patients
Santos, Lívia O.
Vitório, Bianca S.
Branquinha, Marta H.
Pedroso e Silva, Conceição M.
Santos, André L. S.
d'Avila-Levy, Claudia M.
Journal of Antimicrobial Chemotherapy
There is a general lack of effective and non-toxic chemotherapeutic agents for leishmaniasis and there is as yet no study about the effect of HIV peptidase inhibitors (HIV PIs) on Leishmania/HIV-coinfected patients. In the present work, we performed a comparative analysis of the spectrum of action of HIV PIs on different Leishmania spp., including strains obtained from HIV-positive patients receiving or not receiving antiretroviral treatment.
The effects of nelfinavir and saquinavir on Leishmania proliferation were assessed by means of a colorimetric assay (MTT). Subsequently, the effect of nelfinavir on aspartic peptidase activity from Leishmania spp. was assessed by following the degradation of the fluorogenic substrate MCA-G-K-P-I-L-F-F-R-L-K-DNP-Arg-NH2.
Nelfinavir was capable of significantly reducing the multiplication of many Leishmania reference strains and isolates obtained from HIV-positive patients receiving or not receiving antiretroviral treatment. Leishmania major growth was inhibited by ∼50%, while all other flagellates were strongly inhibited (at least 94%), except for a Leishmania chagasi strain obtained from an HIV-positive patient under treatment with highly active antiretroviral therapy (HAART). Culture of this isolate in the presence of nelfinavir induced a considerable reduction in the aspartic peptidase activity. In addition, nelfinavir was also capable of inhibiting the aspartic peptidase activity of all Leishmania strains tested.
The present data contribute to the study of the effect of HIV PIs on Leishmania infection and add new insights into the possibility of exploiting aspartic peptidases as promising targets in order to generate novel medications to treat leishmaniasis.
aspartyl peptidases; Leishmania/HIV coinfection; HIV peptidase inhibitors; chemotherapy; leishmaniasis; proteases
Related Articles
CD8 Cells of Patients with Diffuse Cutaneous Leishmaniasis Display Functional Exhaustion: The Latter Is Reversed, In Vitro, by TLR2 Agonists
Hernández-Ruiz, Joselín
Salaiza-Suazo, Norma
Carrada, Georgina
Escoto, Sofía
Ruiz-Remigio, Adriana
Rosenstein, Yvonne
Zentella, Alejandro
Becker, Ingeborg
PLoS Neglected Tropical Diseases
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 lymphocytes in DCL patients and, additionally, that pre-stimulation with TLR2 ligands can restore the effector mechanisms of CD8 T lymphocytes from DCL patients against Leishmania mexicana-infected macrophages.
Author Summary
Leishmania mexicana causes localized and diffuse cutaneous leishmaniasis. Whereas the former is a benign form the disease, diffuse cutaneous leishmaniasis is a chronic disfiguring disease, for which no cure is available, and the immune cells of these patients respond poorly to the parasite. It has been proposed that the elimination of Leishmania-infected cells by CD8 T cells is crucial for disease control. We compared the functional characteristics of CD8 T cells from patients with localized and diffuse cutaneous leishmaniasis. We found that CD8 T cells from patients with diffuse cutaneous leishmaniasis were functionally exhausted, as compared to patients with the benign form of the disease. We were able to restore functional capacity of these cells by culturing them with molecules that stimulate TLR2. This is the first report showing that stimulation of the TLR2 can restore effector mechanisms in functionally exhausted CD8 cells from patients with diffuse cutaneous leishmaniasis. This finding will help design novel treatment schemes for patients infected with the parasite Leishmania mexicana who have the progressive, incurable form of diffuse cutaneous leishmaniasis.
Related Articles
Cathepsin B-like and cell death in the unicellular human pathogen Leishmania
El-Fadili, A K
Zangger, H
Desponds, C
Gonzalez, I J
Zalila, H
Schaff, C
Ives, A
Masina, S
Mottram, J C
Fasel, N
Cell Death & Disease
In several studies reporting cell death (CD) in lower eukaryotes and in the human protozoan parasite Leishmania, proteolytic activity was revealed using pan-caspase substrates or inhibitors such as carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK). However, most of the lower eukaryotes do not encode caspase(s) but MCA, which differs from caspase(s) in its substrate specificity and cannot be accountable for the recognition of Z-VAD-FMK. In the present study, we were interested in identifying which enzyme was capturing the Z-VAD substrate. We show that heat shock (HS) induces Leishmania CD and leads to the intracellular binding of Z-VAD-FMK. We excluded binding and inhibition of Z-VAD-FMK to Leishmania major metacaspase (LmjMCA), and identified cysteine proteinase C (LmjCPC), a cathepsin B-like (CPC) enzyme, as the Z-VAD-FMK binding enzyme. We confirmed the specific interaction of Z-VAD-FMK with CPC by showing that Z-VAD binding is absent in a Leishmania mexicana strain in which the cpc gene was deleted. We also show that parasites exposed to various stress conditions release CPC into a soluble fraction. Finally, we confirmed the role of CPC in Leishmania CD by showing that, when exposed to the oxidizing agent hydrogen peroxide (H2O2), cpc knockout parasites survived better than wild-type parasites (WT). In conclusion, this study identified CPC as the substrate of Z-VAD-FMK in Leishmania and as a potential additional executioner protease in the CD cascade of Leishmania and possibly in other lower eukaryotes.
cysteine peptidase; cathepsin B; biotin-VAD-FMK; Leishmania; trypanosomes; unicellular organisms
Related Articles
Role of the Trypanosoma brucei natural cysteine peptidase inhibitor ICP in differentiation and virulence
Santos, Camila C
Coombs, Graham H
Lima, Ana Paula C A
Mottram, Jeremy C
Molecular Microbiology
ICP is a chagasin-family natural tight binding inhibitor of Clan CA, family C1 cysteine peptidases (CPs). We investigated the role of ICP in Trypanosoma brucei by generating bloodstream form ICP-deficient mutants (Δicp). A threefold increase in CP activity was detected in lysates of Δicp, which was restored to the levels in wild type parasites by re-expression of the gene in the null mutant. Δicp displayed slower growth in culture and increased resistance to a trypanocidal synthetic CP inhibitor. More efficient exchange of the variant surface glycoprotein (VSG) to procyclin during differentiation from bloodstream to procyclic form was observed in Δicp, a phenotype that was reversed in the presence of synthetic CP inhibitors. Furthermore, we showed that degradation of anti-VSG IgG is abolished when parasites are pretreated with synthetic CP inhibitors, and that parasites lacking ICP degrade IgG more efficiently than wild type. In addition, Δicp reached higher parasitemia than wild type parasites in infected mice, suggesting that ICP modulates parasite infectivity. Taken together, these data suggest that CPs of T. brucei bloodstream form play a role in surface coat exchange during differentiation, in the degradation of internalized IgG and in parasite infectivity, and that their function is regulated by ICP.
Related Articles
Leishmania mexicana Induces Limited Recruitment and Activation of Monocytes and Monocyte-Derived Dendritic Cells Early during Infection
Petritus, Patricia M.
Manzoni-de-Almeida, Daniel
Gimblet, Ciara
Gonzalez Lombana, Claudia
Scott, Phillip
PLoS Neglected Tropical Diseases
While C57BL/6 mice infected in the ear with L. major mount a vigorous Th1 response and resolve their lesions, the Th1 response in C57BL/6 mice infected with L. mexicana is more limited, resulting in chronic, non-healing lesions. The aim of this study was to determine if the limited immune response following infection with L. mexicana is related to a deficiency in the ability of monocyte-derived dendritic cells (mo-DCs) to prime a sufficient Th1 response. To address this issue we compared the early immune response following L. mexicana infection with that seen in L. major infected mice. Our data show that fewer monocytes are recruited to the lesions of L. mexicana infected mice as compared to mice infected with L. major. Moreover, monocytes that differentiate into mo-DCs in L. mexicana lesions produced less iNOS and migrated less efficiently to the draining lymph node as compared to those from L. major infected mice. Treatment of L. mexicana infected mice with α-IL-10R antibody resulted in increased recruitment of monocytes to the lesion along with greater production of IFN-γ and iNOS. Additionally, injection of DCs into the ear at the time of infection with L. mexicana also led to a more robust Th1 response. Taken together, these data suggest that during L. mexicana infection reduced recruitment, activation and subsequent migration of monocytes and mo-DCs to the draining lymph nodes may result in the insufficient priming of a Th1 response.
Author Summary
Leishmaniasis, caused by protozoan parasites belonging to the genus, Leishmania, exhibits clinical symptoms ranging from mild cutaneous lesions to more severe cutaneous or visceral disease. Here, we focus on L. major and L. mexicana, two species that lead to self-resolving and chronic cutaneous lesions, respectively. A strong Th1 response is necessary for resolution of disease following L. major infection. However, L. mexicana infection induces a limited Th1 response resulting in chronic disease. Monocyte-derived dendritic cells are believed to be important in priming the Th1 response during L. major infection, and therefore in this study we evaluated whether there are quantitative and/or qualitative differences in monocyte-derived dendritic cells following L. mexicana infection. We found that fewer monocytes were recruited to the lesions of L. mexicana infected mice as compared to mice infected with L. major. In addition, there were fewer iNOS producing monocyte-derived dendritic cells in the lesions of L. mexicana infected mice and less migration of monocyte-derived dendritic cells to the draining lymph node. Manipulations that allow for increased monocytes in the lesions of L. mexicana infected mice also resulted in a more robust Th1 response. Thus, these findings provide a mechanistic basis for the limited Th1 response observed during L. mexicana infection and also offer a better understanding of the important role that monocytes play during infection with Leishmania.
Related Articles
Leishmania (L.) mexicana Infected Bats in Mexico: Novel Potential Reservoirs
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
PLoS Neglected Tropical Diseases
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.
Author Summary
Leishmaniasis is endemic in southeastern Mexico, where Leishmania (L.) mexicana is the principle parasite species causing disease in humans. Previous studies using methodology for inferring potential biotic interactions between species had shown a high prevalence of bat species as potential hosts for Leishmania, yet bats have not been identified as hosts in Mexico. Motivated by these predictions, we currently analyze whether bats are infected with Leishmania (L.) mexicana and to what extent they serve as hosts for this parasite. We analyzed 420 bats from six states, and through PCR analysis of skin, heart, liver and/or spleen we found that 41 bats (9.77%) of 13 different bat species were infected. Most of the infected bats were netted in an eco-region characterized by plantations grown beneath shade-providing fruit trees that favors breeding conditions for the sand fly vectors and brings them in close proximity with frugivorous, insectivorous or nectarivorous bats. None of the infected bat tissues showed macroscopic lesions. Through artificial infections, we were also able to show that Leishmania (L.) mexicana remains viable and infective after passage through one species of bat. We conclude that various species of bats belonging to the family Phyllostomidae are possible reservoir hosts of Leishmania (L.) mexicana.
Related Articles
Overview of the organization of protease genes in the genome of Leishmania spp
Silva-Almeida, Mariana
Souza-Silva, Franklin
Pereira, Bernardo Acácio Santini
Ribeiro-Guimarães, Michelle Lopes
Alves, Carlos Roberto
Parasites & Vectors
The genus Leishmania includes protozoan parasites that are able to infect an array of phlebotomine and vertebrate species. Proteases are related to the capacity of these parasites to infect and survive in their hosts and are therefore classified as virulence factors.
By analyzing protease genes annotated in the genomes of four Leishmania spp [Leishmania (Leishmania) infantum, L. (L.) major, L. (L.) mexicana and L. (Viannia) braziliensis], these genes were found on every chromosome of these protozoa. Four protease classes were studied: metallo-, serine, cysteine and aspartic proteases. Metalloprotease genes predominate in the L. (V.) braziliensis genome, while in the other three species studied, cysteine protease genes prevail. Notably, cysteine and serine protease genes were found to be very abundant, as they were found on all chromosomes of the four studied species. In contrast, only three aspartic protease genes could be detected in these four species. Regarding gene conservation, a higher number of conserved alleles was observed for cysteine proteases (42 alleles), followed by metalloproteases (35 alleles) and serine proteases (15 alleles).
The present study highlights substantial differences in the organization of protease genes among L. (L.) infantum, L. (L.) major, L. (L.) mexicana and L. (V.) braziliensis. We observed significant distinctions in many protease features, such as occurrence, quantity and conservation. These data indicate a great diversity of protease genes among Leishmania species, an aspect that may be related to their adaptations to the peculiarities of each microenvironment they inhabit, such as the gut of phlebotomines and the immune cells of vertebrate hosts.
Electronic supplementary material
The online version of this article (doi:10.1186/1756-3305-7-387) contains supplementary material, which is available to authorized users.
Leishmania; Leishmania (Viannia) braziliensis; Leishmania (Leishmania) infantum; Leishmania (Leishmania) major; Leishmania (Leishmania) mexicana; Proteases
Related Articles
Regulated Degradation of an Endoplasmic Reticulum Membrane Protein in a Tubular Lysosome in Leishmania mexicana
Mullin, Kylie A.
Foth, Bernardo J.
Ilgoutz, Steven C.
Callaghan, Judy M.
Zawadzki, Jody L.
McFadden, Geoffrey I.
McConville, Malcolm J.
Molecular Biology of the Cell
The cell surface of the human parasite Leishmania mexicana is coated with glycosylphosphatidylinositol (GPI)-anchored macromolecules and free GPI glycolipids. We have investigated the intracellular trafficking of green fluorescent protein- and hemagglutinin-tagged forms of dolichol-phosphate-mannose synthase (DPMS), a key enzyme in GPI biosynthesis in L. mexicana promastigotes. These functionally active chimeras are found in the same subcompartment of the endoplasmic reticulum (ER) as endogenous DPMS but are degraded as logarithmically growing promastigotes reach stationary phase, coincident with the down-regulation of endogenous DPMS activity and GPI biosynthesis in these cells. We provide evidence that these chimeras are constitutively transported to and degraded in a novel multivesicular tubule (MVT) lysosome. This organelle is a terminal lysosome, which is labeled with the endocytic marker FM 4-64, contains lysosomal cysteine and serine proteases and is disrupted by lysomorphotropic agents. Electron microscopy and subcellular fractionation studies suggest that the DPMS chimeras are transported from the ER to the lumen of the MVT via the Golgi apparatus and a population of 200-nm multivesicular bodies. In contrast, soluble ER proteins are not detectably transported to the MVT lysosome in either log or stationary phase promastigotes. Finally, the increased degradation of the DPMS chimeras in stationary phase promastigotes coincides with an increase in the lytic capacity of the MVT lysosome and changes in the morphology of this organelle. We conclude that lysosomal degradation of DPMS may be important in regulating the cellular levels of this enzyme and the stage-dependent biosynthesis of the major surface glycolipids of these parasites.
Related Articles
Hemoglobin digestion in Blood-Feeding Ticks: Mapping a Multi-Peptidase Pathway by Functional Proteomics
Horn, Martin
Nussbaumerová, Martina
Šanda, Miloslav
Kovářová, Zuzana
Srba, Jindřich
Franta, Zdeněk
Sojka, Daniel
Bogyo, Matthew
Caffrey, Conor R.
Kopáček, Petr
Mareš, Michael
Chemistry & biology
Hemoglobin digestion is an essential process for blood-feeding parasites. Using chemical tools, we deconvoluted the intracellular hemoglobinolytic cascade in the tick Ixodes ricinus, a vector of Lyme disease and tick-borne encephalitis. In tick gut tissue, a network of peptidases was demonstrated through imaging with specific activity-based probes and activity profiling with peptidic substrates/inhibitors. This peptidase network is induced upon blood feeding and degrades hemoglobin at acidic pH. Selective inhibitors were applied to dissect the roles of the individual peptidases and determine the peptidase-specific cleavage map of the hemoglobin molecule. The degradation pathway is initiated by endopeptidases of aspartic and cysteine class (cathepsin D supported by cathepsin L and legumain) and continued by cysteine amino- and carboxy-dipeptidases (cathepsins C and B). The identified enzymes are potential targets to developing novel anti-tick vaccines.
Related Articles
Synthetic glycovaccine protects against the bite of Leishmania-infected sand flies
Rogers, Matthew E.
Sizova, Olga V.
Ferguson, Michael A.J.
Nikolaev, Andrei V.
Bates, Paul A.
The Journal of infectious diseases
Leishmaniasis is a vector-borne disease transmitted to human and other mammalian hosts by sand fly bite. Here we show that immunization with Leishmania mexicana promastigote secretory gel (PSG) or a chemically defined synthetic glycovaccine containing the glycans found in L. mexicana PSG can both provide significant protection against challenge by the bite of infected sand flies. Only the glycan from L. mexicana was protective, those found in other species did not protect against L. mexicana infection. Further, neither PSG nor the glycovaccine protected against artificial needle challenge, which is traditionally used in antileishmanial vaccine development. Conversely, an antigen preparation that was effective against needle challenge offered no protection against sand fly bite. These findings provide a new target for Leishmania vaccine development and demonstrate the critical role of the vector in the evaluation of candidate vaccines for leishmaniasis and other vector-borne diseases.
leishmaniasis; vaccine; sand fly; promastigote secretory gel
Related Articles
Naturally azole-resistant Leishmania braziliensis promastigotes are rendered susceptible in the presence of terbinafine: comparative study with azole-susceptible Leishmania mexicana promastigotes.
Rangel, H
Dagger, F
Hernandez, A
Liendo, A
Urbina, J A
Antimicrobial Agents and Chemotherapy
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.
Related Articles
Results 1-25 (311775)
« Previous
Go to page number:
Next »
Remove citation from clipboard
Add citation to clipboard
This will clear all selections from your clipboard. Do you wish proceed?
Clipboard is full! Please remove an item and try again.
Contact Us
PubMed Central Canada is a service of the
Canadian Institutes of Health Research
(CIHR) working in partnership with the National Research Council's
national science library
in cooperation with the
National Center for Biotechnology Information
at the
U.S. National Library of Medicine
(NCBI/NLM). It includes content provided to the
PubMed Central International archive
by participating publishers.


text:
The Afghan "Love Doctor"
Afghan chief Malek Nematullah Halimi has spent more than 20 years breaking down taboos by helping couples with problems in the bedroom. Even though he's not a qualified therapist, his counselling services have attracted couples from all over Afghanistan.
Matthew Bannister also talks to two men from rival gangs in Birmingham, UK, who risked their lives to agree a truce between the opposing sides.
An Inuit campaigner tells why she believes seal hunting is essential to her community's survival.
And we meet the Malian man who tried to save ancient manuscripts from Islamist militants.
Picture: Malek Nematullah Halimi, the Afghan "Love Doctor".
Last on
More episodes
Aaju Peter
Aaju Peters is a campaigner for the rights of her Inuit people of the Canadian Arctic, and she works to keep their ancient traditions alive.

subset: commoncrawl


text:
Sunblock Lotion - Baby Continuous Protection
Active: Avobenzone 3%...Purpose: Sunscreen, Homosalate 10%...Purpose: Sunscreen, Octisalate 5%...Purpose: Sunscreen, Octocrylene 2.8%...Purpose: Sunscreen, Oxybenzone 6%...Purpose: Sunscreen. Inactive: Water, Butyloctyl Salicylate, Styrene/Acrylates Copolymer, Silica, Diethylhexyl 2, 6-Naphthalate, VP/Hexadecene Copolymer, Dimethicone, Phenoxyethanol, Avena Sativa (Oat) Kernel Flour, Ethylhexylglycerin, Caprylyl Methicone, Glyceryl Stearate, PEG-100 Stearate, Trimethylsiloxysilicate, Sodium Polyacrylate, Ethylhexyl Stearate, Methylparaben, Behenyl Alcohol, Xanthan Gum, Propylparaben, Ethylparaben, Disodium EDTA, BHT, Trideceth-6.
Directions: Apply generously 15 minutes before sun exposure. Reapply after swimming or sweating, immediately after towel drying, or at least every 2 hours. Sun-Protection Measures: Spending time in the sin increases your risk of skin cancer and early skin aging. To decrease this risk regularly use a sunscreen with a Broad Spectrum SPF value of 15 or higher and other sun-protection measures including: Limit time in the sun, especially from 10am-2pm, or wear long-sleeved shirts, pants, hats, and sunglasses. Children under 6 months of age: Ask a doctor. Other Information: Protect this product from excessive heat and direct sun. May stain some fabrics.
Other Info
Johnson & Johnson Consumer Products Company
Skillman, NJ 08558-9418
Safety Warnings
For external use only. Do not use: on damaged or broken skin. When using this product: keep out of eyes. Rinse with water to remove. Stop use and ask a doctor: if rash occurs. Keep out of reach of children. If swallowed, get medical help or contact a Poison Control Center right away.


text:
Standing out in your industry is more important than ever before, and thought leadership is one of the most effective ways of you can rise above the competition.
PR Insiders
How to Use Thought Leadership to Energize Your BrandJune 5th, 2015 by Ronn Torossian
7 PR Tips Not (Easily) Found In Your AP Style GuideMay 28th, 2015 by Shawn Paul Wood
PR pros on a quest toward communications supremacy would do well to check out some of these hidden gems from the AP Style Guide.
How to Bring Life to Shareable ContentMay 26th, 2015 by Alex Clough
Creating shareable content for the modern audience is a challenge that can be overcome by following these key principles.
Making Time for Face Time With PR ClientsMay 18th, 2015 by Jennifer Leckstrom
While it's possible to conduct business without ever meeting in-person with clients in certain cases, but we shouldn't forget about the value of such interactions.
3 Reasons Why Email Marketing And PR Agencies Should Work TogetherMay 14th, 2015 by Christopher Lester
PR practitioners are looking for ways to meld modern marketing tactics into their plans. And many are discovering that email marketing is one of the best crossover tools.
PR Insider: How Legacy Brands Adapt to a New AgeMay 7th, 2015 by Kathleen Delaney
Legacy brands face a unique challenge in maintaining relevancy in the digital age. Here is how one such company met and overcame that challenge.
PR Insider: Handling Communications During a Product RecallMay 4th, 2015 by Todd Harris
Preparation is crucial in communicating with the public, the media, and employees about a product recall.
PR Insider: Making Contacts in the Newspaper NewsroomApril 27th, 2015 by Andrew Blum
Many newspaper newsrooms these days still have a beat system, but there are also web sites, video and a constantly changing media landscape to contend with.
PR Insider: Making Facebook Work for PRApril 23rd, 2015 by Shawn Wood
PR is about presence, positioning, and promotion. Facebook can help your client accomplish all three, if you use the platform for the reason it was created.
PR Insider: Keeping It Short and Simple With the MediaApril 20th, 2015 by David Hamilton
For PR professionals, especially those tasked with pitching story ideas to the media, the mantra to remember is "Keep It Short and Simple."


text:
Hotels in Haggatt Hall, Barbados
Search & Compare Haggatt Hall Hotels
Trending now in Haggatt Hall
- Most bookedAccra Beach Hotel & Spa
179 people looked at this hotel in the last hour
Most recent review
"Stayed 4 days/3 nights in early June. Hotel staff was very accommodating, polite and responsive. Was our first stay in Barbados but we thought the location of Accra was really good - walking distance to some fantastic restaurants, handy little store in the mall across the street, etc."
C, Jun 2016, US
32 people looked at this hotel in the last hour
Most recent review
"When we arrive , the reception was incredible, the hotel staff is wonderful and helpful, rhum punch, made us relax and the hall is very pretty, the appartement is incredible kitchenette and balcony to overlook the sea. The restaurant is very good and Irvin is the best barman. The pool is relaxing..."
A Traveller, Apr 2016, CA
23 people looked at this hotel in the last hour
Most recent review
"We had a great stay here. The facility is well maintained, rooms clean, and staff friendly. The majority of resort occupants are/were time-share owners and are long-time visitors. It definitely has a 'neighbourhood' vibe. The beach is about 10-12min walk (but beware there are rarely sidewalks in ..."
A Traveller, Feb 2016, CA
22 people looked at this hotel in the last hour
Most recent review
"My 3 relatives and I came to Barbados for a family occasion and are very happy to have decided on this house after several hours of searching for suitable accommodation. Zen Break is perfectly decorated. The furniture is modern and very comfortable. And in Barbados, what nailed it, was the fact t..."
A Traveller, May 2015, IE
12 people looked at this hotel in the last hour
Most recent review
"Great service; Greg was very flexible and a wonderful host who graciously accommodated our last-minute requests."
A Traveller, Nov 2015, US
11 people looked at this hotel in the last hour
Most recent review
"Beautiful hotel with very attentive staff - we will go back and stay longer. Breakfast was creative, beautifully presented and made with high quality fresh ingredients."
A Traveller, Mar 2016, GB
10 people looked at this hotel in the last hour
9 people looked at this hotel in the last hour
Most recent review
"Hotel was clean. A/C works but not that good and room bedroom light was not working"
s, Apr 2016, US
5 people looked at this hotel in the last hour
Most recent review
"I really enjoyed Southern Surf Beach Apartments. Our room was spotless and comfortable. The air conditioner worked very well. I really liked having a kitchen and the extra space. I was travelling with a friend for the Barbados Swim Festival, and we wanted a place to keep and prepare "healthy" sna..."
A Traveller, Nov 2015, US
5 people looked at this hotel in the last hour
Most recent review
"Christy's house is a haven of alternative living away from the beach resort world. Homely furnishings and an abundance of plants give an artistic, bohemian ambience. Christy will meet you at the airport and is on hand to organise trips, services, taxis etc and her breakfasts are delicious - fresh..."
A Traveller, Jun 2014, GB
Best hotels in Haggatt Hall
If you are looking for Haggatt Hall hotels in a particular area, you can use the search feature at the top of the page to search by neighborhood, landmark, address or zip code, or you can search by using the built-in map feature. You can also browse hotels in Haggatt Hall by type, amenities, and star-rating so that you can find the best hotel. Currently, top rated hotels in Haggatt Hall are:
- Malfranza Apartments
- ZenBreak - Anidale Spacious Condo
- Sweetfield Manor Historic Inn and B&B
- South Beach Hotel
- The Barbados CHI Centre
The location of your Haggatt Hall hotel is important. If you are traveling to Haggatt Hall, &StateorCountry for business, you may want to consider getting a hotel close to the airport or near your meeting venue. By doing so, you can really cut down on transportation costs and time. You may also want to book a hotel that offers free wifi just in case you need to get some work done.
If you are traveling for vacation, figure out which attractions or landmarks you want to see and book a hotel that is within close proximity. Although hotels near attractions might cost a little more, you may find that spending those extra few dollars is worth the convenience. You can even find hotels that align with your interests. For example, if you are looking for a relaxing vacation, a spa hotel may be the perfect choice for you.
You'll also want to determine which type of accommodations will suit your needs before booking a hotel in Haggatt Hall. If you're traveling with the kiddos, a family friendly hotel will offer larger rooms and more sleeping arrangements. For those traveling for an extended period of time, booking a hotel suite with a kitchenette is a smart move and will feel a little more like home. If you are budget conscience and just need a place to rest your head, a hostel or motel may be all you need.
If you are looking to score the best discounts and deals, use the filter to sort by price from low to high. You can book a hotel in Haggatt Hall for as low as &PricesFrom per night. If you are looking to cut down on expenses, look for hotels that include free breakfast and parking. Some hotels even offer free shuttle service to and from nearby places. You can also sort by star rating, proximity, and guest rating.
Before you book, check out user reviews. Hotels.com has 0 reviews in Haggatt Hall. We show reviews from Expedia and TripAdvisor so you know that you are reading genuine, reliable reviews. Be sure to download the Hotels.com app so you can book your reservations on the go, view amenities, and save the hotels you want to view and compare later.


text:
where can I get the prospectus (on line) of the high priced preferreds RBS-pH and rbs-pF?
It has been quite some time, but I looked at these on the SEC website, using Quantum Online to get the date of inception then using the SEC site to find the prospectus.
The H and F prospectus are virtually identical in form, language, and conditions as the the L preferred (prospectus is linked to at Quantum Online site). I say this based on memory, but I was pretty focused on these a back after the collapse in 2008-09.


text:
While on a golf weekend with a couple of friends we decided to try Kaminski Brothers Chop House after a long day of golf. We were staying at the Chula Vista Resort and since it is right on the resort grounds we didn't have too far to go! We went on a Saturday night around 6:00 p.m. without reservations but we were seated right away (probably because it was off season). The menu was a simple one page collection of appetizers, steaks, entrees and sides. We decided to start with the calamari for our appetizer. It came mixed with peppers and had a sauce on the side. It was great, but as we soon came to find out, it may have been too much for us! Each entree comes with their Garbage Salad. It was a weird name for such a great salad! The waiter brings out a cart with a bowl of pre-mixed field greens, eight bowls of other items that can be mixed in such as red onions, eggs, bacon, baby shrimp, tomatoes and three others (sorry, cannot remember what they were!) and a creamy caesar dressing. It was delicious! This course comes served with their bread, which was o.k.
We ordered a side of mashed potatoes to be served with our entree. All of the sides are served family style. The mashed potatoes were very good, but it was enough for more than 10 people (there were only three of us!). I had the scallops wrapped in bacon and my friends had filet mignon and the petite filet. It was all excellent tasing!
Be prepared to spend quite a bit of money especially if you order beer or wine. While I would not have this on a regular basis, for a one time meal Kaminski Brothers Chop House is an excellent choice especially if you like beef!
Own or manage this property? Claim your listing for free to respond to reviews, update your profile and much more.


text:
The latest round of Iron Builder has begun!
The Battle Begins!
In the coming weeks Rod and Pascal will post their various MOCs below.
Iron Builder is a contest sponsored by the Builder's Lounge group on Flickr. In the past, it has been more of a private contest ( though many many of the excellent entries have seen the light via Brother's Brick and other popular LEGO blogs ) and we have decided to more readily share the MOCs with non Lounge members by posting them as a common thread for everyone to enjoy.
The basic rules of the contest require that the two builder's receive a batch of the 'secret ingredient' part. In this case, the Blue Torso Mechanical - Battle Droid
The two builders have one month to produce as many high end builds as possible to submit to the panel of judges.
The winner of the contest becomes ( or remains ) one of the FOUR Iron Builders.
Current IBs are:
Olog ( Recently defended against Legoloverman )
2MuchCaffeine ( recently defended against Lino )
Bruceywan ( recently dethroned Iron Buider Legohaulic )
Tadashistate ( recently defended against MonsterBrick )
Combatants fight for bragging rights and , more importantly, the winner is built a trophy by the conquered!
Feel free to cheer for your favorite builder , encouragement is encouraged!
Wibbly wobbly timey wimey.
Mech me this, mech me that...
Oops! Forgot to post this here previously...
The showing has been excellent so far.
One week left to go in the battle , last day for entries is the 30th.
Best of luck to both of our competitors!
Feruss/Bilda Robotics Company - Factory No9
And the video of it in action...
The Results are in!
For our judges this round, I wanted some seasoned experienced builders. ( 2 ) of them are former IBs and one of them likes to spell his name backwards.
Please welcome Legohaulic, SlyOwl and SirNadroj!
Here's what they had to say about our handsome and daring contestants in what has been called the "Battle of the Vignettes" and "Aren't these both the same guy" ?
Judge #1
Building skill: 18/20
Presentation: 20/20
Use of mystery part: 16/20
Total: 54/60
Building skill: 17/20
Presentation: 20/20
Use of mystery part: 18/20
Total: 55/60
Well...I'm still not convinced that these two builders aren't the same person, but as far as online personas, Pasukara wins but only slightly. I had a really tough time deciding between the two. Both builders have almost identical styles of building and presentation. I really can't tell the difference between them on most of the MOCs. It took me a while to decide which builder should win and here's my explanation or rationalization for why.
When it comes to building skill, I believe that Rod has a slight advantage. I feel that his models are more aesthetically pleasing and fun to look at. I also feel like there was a bit more time invested in most of his models.
Presentation is a tie between the two. Both photograph their MOCs in almost the exact same way and it looks outstanding.
Lastly, in regards to the use of the mystery part, I feel that Pascal wins here. While both builders used the part exclusively as mechanical bits, Pascal seemed to use the part in a more diverse way. Vacuums, conveyor belts, park benches, treads, and finally molecules have won me over in their variety. In my opinion, I feel that diversity of part usage holds more weight when tallying the final score.
Congrats both of you on a battle well fought. I think we shall all remember the battle of the vignettes regardless of the victor.
Judge #2
Building skill: 16/20
Presentation: 16/20
Use of mystery part: 17/20
Total: 49/60
Building skill: 17/20
Presentation: 17/20
Use of mystery part: 14/20
Total: 48/60
This, in my opinion, is one of the closest Iron Builder competitions I've seen. All-around impressive entries from both sides, and it ended up being the little things that helped me score each contestant the way I did. Both of your models are very fun and very playful. Again, quality-wise both contestants produced comparable models - it seems that pasukaru76's models were just *slightly* more complex technique-wise, which is why I scored that category 17-16 pasukaru76. Presentation was also borderline perfect on both sides, but the variety of pasukaru76's photos made me score it 17-16 pasukaru76 again. Lastly was the use of mystery part, and this is what tipped the scale in 2muchcaffeine's favor. I feel 2muchcaffeine used the mystery part a bit more prominently in his models than pasukaru76, and for me that is almost more important in this type of competition than the other two categories, so I had to score it 17-14 2muchcaffeine, which tips the scale in 2muchcaffeine's favor 49-48. Well done all around gents!
Judge #3
Building skill: 17/20
Presentation: 18/20
Use of mystery part: 16/20
Total: 51/60
Building skill: 18/20
Presentation: 17/20
Use of mystery part: 18/20
Total: 53/60
Rod's use of the mystery part was really good - though evidently it does tend to lend itself to small vehicles! These on their own wouldn't have done so well, but coupled with the backdrops and thematic series, bumped them up to the next level. I thought the Boonta Eve Podrace was fantastic.
Pascal's use was a bit more varied than Rod's, I thought - the cable car, the molecule and the small minifig scenes in particular pushed it ahead. As lots of people have said, it was tricky to know who was building what! However, I do think Chris' comment is worth noting - "What you have is awesome. but it needs more context."
Certainly, it's a difficult part to use in blue - so kudos to both builders for getting such a range of models out of it. Great fun to watch.
A close 2-1 split decision means.........
Our winner is ....... Pascal!
A new Iron Builder is born!
Well done Pascal and 2MC, some really cool builds along the journey!
Here is the All Iron Builder flickr group
Perhaps you will discover a past battle of bricks and blood that you missed.
Feel free to look around and admire the Mad sKILLz.
: )


text:
Opening times for Abeno
Opening and closing times for Abeno
Today (Sunday) Abeno opens at 12noon and closes at 10pm
Location map of Abeno
Underground Stations
Railway Stations
Car Parks
Hotels Near
What's On Today
Near Abeno
Charlie and the Chocolate Factory
Theatre | Until 7th January 2017
Theatre Royal Drury Lane 3 minutes walk from Abeno
Nominated for a staggering seven gongs in the 2014 Olivier Awards including Best New Musical, Charlie and the Chocolate Factory ...More
Matilda The Musical
Theatre | Until 28th May 2017
Cambridge Theatre 3 minutes walk from Abeno
The Royal Shakespeare Company's multiple award-winning musical adaptation of Roald Dahl's celebrated children's book Matilda is a big hit at ...More
The Lion King
Theatre | Until 4th June 2017
The Lyceum Theatre 3 minutes walk from Abeno
The Lion King tells the story of Simba - the epic adventure of his battles, his friendships, his dreams and ...More
Theatre | Until 8th January 2017
Ambassadors Theatre 4 minutes walk from Abeno
Stomp stages an aural assault on your senses at the Ambassadors Theatre. There is no plot, no dialogue and nobody ...More
DJ Lady Lloyd
Clubbing and Nightlife | Until 18th December 2016
Ku Bar 6 minutes walk from Abeno
Retro pop classics from the 1980s and 1990s. With support from DJ Vicki Vivacious....More
Daydreaming with Stanley Kubrick
Exhibitions | Until 24th August 2016
Somerset House 6 minutes walk from Abeno
In a unique summer exhibition, Somerset House explores the impact of one of the most innovative and influential filmmakers of ...More
Strangeness In Space In Conversation With Robert Ross
Exhibitions | On 31st July 2016
Leicester Square Theatre 6 minutes walk from Abeno
Trev & Simon, Sophie Aldred and Clare Eden talk to Ross about making the space-based radio series....More
Stage Against The Machine
Exhibitions | On 31st July 2016
Leicester Square Theatre 6 minutes walk from Abeno
Suzy Wilde, Dave Black and Simone Belshaw take speak frankly about societal and personal issues....More
Magic Carpet Storytelling (Under 5s)
Kids Events | Until 31st July 2016
National Gallery 6 minutes walk from Abeno
Stories inspired by pictures in the National Gallery....More
Sunday Jazz Lunch!! Richard Shelton With The Renato D'aiello Quartet
Concerts | On 31st July 2016
Ronnie Scott's 7 minutes walk from Abeno
The vocalist, accompanied by the saxophonist and his group, performs smooth jazz and easy listening numbers....More
London Hotels
From cheap budget accommodation to luxury 5 star suites see our special offers for Abeno hotels, including Hotel Indigo London Kensington - Earls Court from 72% off.


text:
7-Piece Ceramic Cookware Set£25 instead of £70.96 for a 7-piece ceramic cookware set inc. saucepan, frying pan and saucepots in red, black or cream from Wowcher Direct - save 67%
Save 65%
Time left to purchase
- Get a stylish and durable set of ceramic cookware.
- 16cm saucepan, 20cm and 24cm saucepots and 24cm frying pan.
- Plus lids for the saucepan and saucepots.
- Choice of black, red or cream.
- Professional dishwasher-safe quality in non-stick aluminium.
The Fine Print
- May buy multiples.
- Voucher valid until 16th Mar 2014.
- Delivery costs an additional £3.99 per set.
- Please allow up to 10 working days for delivery after redeeming.
- Subject to availability.
Full Details
Whilst a workman should never blame their tools for a poor job, it's perfectly acceptable for a home cook to blame dodgy pans for rubbish food. Swap yours for today's deal...
We're offering you a 7-piece ceramic cookware set in red, cream or black for just £25, which compares to direct2public's price of £70.96 (correct as of 26.2.14).
Whether it's the obligatory Sunday fry-up or a dinner party for all your closest friends, no domestic goddess' kitchen is complete without a set of professional quality non-stick aluminium pans - so it's lucky we're here to stock your kitchen cupboards with a matching 7-piece set! Each pan is dishwasher-safe and suitable for use on most hobs. Simply choose whether you'd prefer a red, black or cream set that includes the following:
- 1x 16cm saucepan
- 1x 20cm saucepot
- 1x 24cm saucepot
- 1x 24cm frying pan
- 3x saucepan and saucepot lids
How to Redeem
- When the deal ends we'll send you your unique wowcher code.
- Head to the Wowcher redemption page using the link we send you.
- Select your chosen colour.
- Enter your wowcher code and delivery details.
Once purchased, you can redeem this wowcher towards another wowcher or claim a Wowcher Wallet Credit. Click here for more info.
Health MOT - Over 30 Locations!£79 instead of £349 for a Wellwoman or Wellman health MOT with UK Health Screening - choose from over 30 locations and save 77%
Save 77%
Time left to purchase
4nt Pontins Summer Family Break for 4 - 4 Coastal Locations! From £99 (at Pontins) for a four-night summer family break for up to four people - save up to 29%
Bookkeeping, HR & Payroll Course - Online or Classroom Options!£16 instead of £199 for a certified online HR, bookkeeping and payroll course or from £405 for a classroom course with Mary Jay Kenny - save up to 92%
Save 92%
Time left to purchase


text:
Fibersym® RW Resistant Wheat Starch is a convenient and rich source of dietary fiber that can be formulated in a wide array of foods with minimal processing adjustments. This RS4 starch is ideal for incorporation in foods that emphasize benefits related to health and wellness.
Fibersym RW also acts as a prebiotic and can reduce the caloric content of foods while providing a high source of dietary fiber.


text:
Medium Stainless Steel Sink StrainerProduct Code: SLSN002
Availability: In Stock
Reg. Price $2.50
Our Price: $0.81 Savings $1.69
The Medium Stainless Steel Sink Strainer will help your pipes run clear. This fine mesh strainer holds onto food waste while letting water through. Simply insert the strainer into your sink hole while washing dishes and empty it periodically as it fills with waste.
- Sink Strainer
- Stainless steel construction
- Extra fine mesh lining
- 2-3/4" dia.
- Brand:
- Thunder Group
- Model #:
- Ship weight:
- 1 oz
- Sold As:
- 1


text:
Welcome to Saint George Parish Community!
"I rejoiced when I heard them say let us go to the house of the Lord." Psalm 122
If you are new to the area, just visiting, or looking to learn more about the Catholic faith, we invite you to join us for Mass and one of our many community events. Come and learn more. Or contact us by phone or Email.
Reverend Leo P. Oswald
If you would like to register online, please click on the Newcomers tab for the Registration Form and fill in the information. Once submittted, you will be contacted by a member of our staff to confirm the information and to let you know that it has been processed.
Bible Camp
Bible Camp will be held the first week in August 2016. Registration forms are available at the Rectory Office. We will meet in the Parish Hall from 10:00 AM to 1:00 PM Monday through Thursday and then we will head to Dorney Park Amusement Park and Wild Water Kingdom on Friday. Don't miss out on the fun. Please see the Parish Bulletin for details.
Pope Francis Proclaims "Year of Mercy."
December 8, 2015 to November 20, 2016
The Extraordinary Jubilee Year of Mercy is a remarkable occasion during which the entire Catholic Church, under the humble leadership of Pope Francis, opens wide the doors to the saving mercy of Christ. The Year of Mercy is celebrated from December 8th, 2015 - the Feast of the Immaculate Conception and the 50th anniversary of the closing of Vatican II - to the Solemnity of Christ the King on November 20, 2016.
During this special period of time in the Church, Pope Francis calls all Catholics to be profound witnesses to mercy and to "find the joy rediscovering and rendering fruitful God's mercy, with which we are all called to give comfort to every man and every woman of our time." To learn more, please visit the Vatican's official Year of Mercy website at www.im.va or the United States Conference of Catholic Bishops website at www.usccb.org.
We are especially encouraged to practice the Spiritual and Corporal Works of Mercy, to receive the Sacrament of Penance as often as needed, and to pray the Chaplet of Divine Mercy. Please plan to spend some quiet time throughout this Year of Mercy with the Blessed Sacrament on Fridays, any time between the hours of 8:00 AM and 4:00 PM, and on Thursday evenings at 7:00 PM for the recitation of the Most Holy Rosary and the Divine Mercy Chaplet in St. George Chapel.
Lord Jesus Christ, you have taught us to be merciful like the heavenly Father, and have told us that whoever sees you sees Him. Show us Your face and we will be saved. Your loving gaze freed Zacchaeus and Matthew from being enslaved by money; the adulteress and Magdalene from seeking happiness only in created things; made Peter weep after his betrayal, and assured Paradise to the repentant thief. Let us hear, as if addressed to each one of us, the words that You spoke to the Samaritan woman:"If you knew the gift of God!" You are the visible face of the invisible Father, of the God who manifests His power above all by forgiveness and mercy:let the Church be Your visible face in the world, its Lord risen and glorified. You willed that your ministers would also be clothed in weakness in order that they may feel compassion for those in ignorance and error:let everyone who approaches them feel sought after, loved, and forgiven by God. Send Your Spirit and consecrate every one of us with its anointing, so that the Jubilee of Mercy may be a year of grace from the Lord, and your Church, with renewed enthusiasm, may bring good news to the poor, proclaim liberty to captives and the oppressed, and restore sight to the blind. We ask this through the intercession of Mary, Mother of Mercy, You who live and reign with the Father and the Holy Spirit for ever and ever. Amen.
Remembering Father Denny
Saturday, July 16, 2016
4:30 PM St. George Church
On Saturday, July 16th we will be celebrating a Memorial Mass for Reverend John T. Denny, OSA in St. George Church. All are welcome to join us as we remember Father Denny and pray for him and his family.
Our Mass Schedule
Saturday: 4:30 PM
Sunday: 8:00 AM and 10:00 AM
Weekdays: Mon-Wed-Fri 4:00 PM, Tues-Thurs-Sat. 8:00 AM
Confession: 3:30-4:15 PM Every Saturday at St. George Church and Mondays during Lent and Advent following the 4:00 PM Mass. Also by appointment.
Our Rectory Hours
Monday thru Friday: 9:00 AM to 5:00 PM
Saturday: 9:00 AM-3:00 PM
Sunday: 8:30 AM-12:30 AM
Casserole Weekend
Empty casserole pans and suggested casserole recipes are placed at the main (front) doors of the church on the last weekend of every month. Filled, frozen casseroles are then returned on Casserole Weekend, which falls on the first weekend of each month unless otherwise announced. Casseroles can be returned directly to the Rectory Office on Saturday between the hours of 9:00 AM and 3:00 PM, and on Sunday between 8:30 AM and 12:30 PM. Casseroles can also be dropped off Monday thru Friday between the hours of 9:00 AM and 5:00 PM. All Casseroles are picked up by and delivered to St. John's Hospice in Philadelphia where meals are provided for the homeless. Please join us in our efforts to end hunger. God bless you for reaching out to those in need of a meal.
Ask George
Question: Why do we address priests as "Father" when it clearly states in Matthew 23:9, "Call no one on earth your Father; you have but one Father in heaven."
Answer: Perhaps the most pointed New Testament reference to the theology of the spiritual fatherhood of priests is Paul's statement, "I do not write this to make you ashamed, but to admonish you as my beloved children. For though you have countless guides in Christ, you do not have many fathers. For I became your father in Christ Jesus through the gospel" (1 Cor. 4:14–15).
Peter followed the same custom, referring to Mark as his son: "She who is at Babylon, who is likewise chosen, sends you greetings; and so does my son Mark" (1 Pet. 5:13). The apostles sometimes referred to entire churches under their care as their children. Paul writes, "Here for the third time I am ready to come to you. And I will not be a burden, for I seek not what is yours but you; for children ought not to lay up for their parents, but parents for their children" (2 Cor. 12:14); and, "My little children, with whom I am again in travail until Christ be formed in you!" (Gal. 4:19).
John said, "My little children, I am writing this to you so that you may not sin; but if any one does sin, we have an advocate with the Father, Jesus Christ the righteous" (1 John 2:1); "No greater joy can I have than this, to hear that my children follow the truth" (3 John 4). In fact, John also addresses men in his congregations as "fathers" (1 John 2:13–14).
By referring to these people as their spiritual sons and spiritual children, Peter, Paul, and John imply their own roles as spiritual fathers. Since the Bible frequently speaks of this spiritual fatherhood, we Catholics acknowledge it and follow the custom of the apostles by calling priests "father." Failure to acknowledge this is a failure to recognize and honor a great gift God has bestowed on the Church: the spiritual fatherhood of the priesthood. Catholics know that as members of a parish, they have been committed to a priest's spiritual care, thus they have great filial affection for priests and call them "father." Priests, in turn, follow the apostles' biblical example by referring to members of their flock as "my son" or "my child" (cf. Gal. 4:19; 1 Tim. 1:18; 2 Tim. 2:1; Philem. 10; 1 Pet. 5:13; 1 John 2:1; 3 John 4).
All of these passages were written under the inspiration of the Holy Spirit, and they express the infallibly recorded truth that Christ's ministers do have a role as spiritual fathers.
Jesus is not against acknowledging that. It is He who gave these men their role as spiritual fathers, and it is His Holy Spirit who recorded this role for us in the pages of Scripture.
It is also interesting to know that the Pope, in Italian, is referred to as Papa. Jesus refers to His father as Abba, also an intimate term for God as Father. In simple terms, the role of a priest is to be a shepherd/father to his flock, who are the "family" of the Church.
We received another interesting question regarding the Our Father prayer, which we will address in another Bulletin issue. Thank you for your questions. We are happy to address your concerns.


text:
Game Details - Parkview HS
The Kickapoo Chiefs football team has an away conference game @ Parkview on Friday, August 19 @ 7p.
MaxPreps is looking for one person to be the Official Scorer! This person will provide LIVE play-by-play for fans to follow along during the game.
If you have any corrections for the Kickapoo football schedule please contact Support.


text:
School of the Arts
School of the Arts News
Racism and Discrimination To Be Explored in Lecture
Harvard University faculty member Tommie Shelby will speak on "Racism, Discrimination, and Equal Opportunity"
Ploughshares Releases Collection of Digital-First Stories
A new volume of Ploughshares Solos has been published.
'Cool' Display for a Chilly Night
The façades of massive apartment buildings and small businesses were covered in projected images, text, and video clips
Emerson College Faculty Honored
Annual event honors faculty members for their service
First Women in Film & Media Summit Slated for November
The first Women in Film & Media Summit aims to create opportunities for women in the industry.
TV Studio Transitions to Latest Technology
The Tufte facility has been transitioned to a platform that is contemporary with professional HD facilities.
Students Discuss Film About Former Gay Activist
An outspoken gay activist makes a 180-degree reversal, rejecting his sexuality, spreading anti-gay opinions, and joining the Christian clergy.
'Suffragette' Essay Contest Launched
Emerson students are invited to enter an essay contest related to the new film Suffragette.


text:
NCAA Football 14: New game has same feel
NCAA Football 14 has a new-game feel and gives couch-potato quarterbacks more control over their coaches and teams. The game has an added element of physics as players deal with true weight and momentum in a fast-paced, hard-hitting game. Loading times are not as long. There are changes to the overall presentation, along with new features.
Yet the game has an old familiar look and feel to it. It's still fun and realistic, but the basic elements haven't changed.
Developer EA Tiburon not only improved the physics and force impact on both sides of the football, but adjusted the running game to include hard cutting, direction changes, improved run blocking and ball carrier mobility. The game has new big hits and stiff arms galore.
Choose any team in the country and run the latest read-option offense, then scramble against gravity to gain a few extra yards.
There's new commentary from ESPN announcers like Rece Davis, Brad Nessler and Kirk Herbstreit. Fan chants have also been added to the overall stadium atmosphere, fight songs and mascot antics.
NCAA Football 14 allows players to build a team from a collection of more than 2,500 licensed former NCAA athletes in "Ultimate Team" mode.
Even so, with all the new flashy features, NCAA Football 14 feels like previous PS3 versions. It must be hard to innovate a game that already has practically everything.
Regardless, the game is still very fun to play. You still feel like you are in the game as you take in the sounds of the field, look over the defense, call an audible and stun the defense with a big play.
I doubt that will never change.
Actually, NCAA Football in its current form may become a collector's item since the NCAA has said it will no longer allow Electronic Arts Inc. to use its logo starting next year. The move ends a lucrative partnership and "comes as the NCAA fights a high-profile lawsuit that says the governing body owes billions of dollars to former players for allowing their likenesses to be used for free," according to the Associated Press.
Game: NCAA Football 14
Platforms: PlayStation 3, Xbox 360
Price: $59.99
ESRB rating: E for everyone
- Rupert Murdoch vows Fox News without Ailes is...
- Fox needs to find a successor to man who...
- Froome crashes in Tour, finishes stage with...
- Warriors' Draymond Green settles assault case...
- Trump takes time from victory lap to swat at...
- Warner Bros. calms fans with fun 'Justice...
- A title and ship for new 'Star Trek' series:...
- New artificial reef is sunk off Florida's...
- Trump takes time from victory lap to... 13
- Rupert Murdoch vows Fox News without... 12
- Stephen Colbert is back to take on the... 9
- Chris Hicks: Movies to watch over the... 3
- Fox needs to find a successor to man... 2
- Police: Munich suspect was obsessed... 1
- Why 'Rogue One' will be... 1
- Life lessons keep coming from Star Trek... 1


text:
Electrical problem
1995 Acura Integra 4 cyl Front Wheel Drive Manual 150000 miles
My acg fuse keeps blowing in the car. It has an engine swap in it and the wiring harness is kinda jacked up. It was like that when I bought it. But I'm wondering if my alternator is bad or maybe if there is an open wire causing a short. Help!
Saturday, November 6th, 2010 AT 7:58 PM


text:
Virtual Families 2: Our Dream House Answers for iPhone - iPadHome / iPhone - iPad /
Game Questions & Answers (Q&A) service lets you ask questions about video games for game consoles or PC games. So ask your Virtual Families 2: Our Dream House question for iPhone - iPad and get answers from real gamers or answer questions and share your insights and experience with the rest of the gaming community.
- Developer: Last Day Of Work
- Publisher: Last Day Of Work
- Genre: Simulation Virtual
- Release: Dec 6, 2012
- ESRB: Not Set
Question & Answers
How do I get more random emails?
I'm new to the game
My first generation peeps would get an email almost every hour for their whole lives. Now I'm on my second generation and I haven't gotten an email other than a marriage proposal and an email to player..what happened to all my mail?? I need more random events to make my game more interesting!
Answer from: Magic
Buy the dice from the special upgrades and there are random event dice . But be warned You can get bad events
Answer from: Virtual Dreamer
When you get those E-mails you don't get a random event. Actually, those E-mails get to be a pain because they are pete and repeat. In the beginning I liked them because I was hoping..LOL. Good luck w/your game.
Answer from: Chunky choclate
E-mails can sometimes be bad to some times they take money away from you there is the one that you have overpaid your taxes gives you money the one where you underpay pay your taxes takes money from you
Answer from: Stupid people
Ever heard of this thing called a period? You used it at the end of a sentence. "Chunky Choclate".
Virtual Families 2: Our Dream House - Gameplay Review Trailer
- Oct 31, 2013, Views: 4301
Virtual Families 2: Our Dream House - Trailer
- Sep 12, 2014, Views: 3183
Virtual Families 2: Our Dream House - Gameplay Video
- Sep 12, 2014, Views: 2157
Virtual Families 2: Our Dream House - Part 2 Gameplay
- Sep 12, 2014, Views: 1971
- Deus Ex: Mankind Divided, (PS4)
- The Journey: Bob's Story, (PC)
- Mafia 3, (PS4)
- Forestry 2017 - The Simulation, (XONE)
- Tayutama 2: You're The Only One, (PC)
- Call Of Duty: Infinite Warfare, (PS4)
- God Eater Resurrection, (PC)
- Red Comrades 2: For The Great Justice, (PC)
- Mystery Of Rivenhallows, (PC)
- Road Not Taken, (iOS)
- Cubway, (iOS)
- Colorcube, (iOS)
- Doodle God Griddlers, (iOS)
- Crusader Kings: Chronicles, (iOS)
- Roads Of Rome: New Generation, (iOS)
- Kubic, (iOS)
- Guild Of Dungeoneering, (iOS)
- Dead Shell: Roguelike RPG, (iOS)
- Quell Zen, (iOS)
- Lanota, (iOS)
- Maquis Board Game, (iOS)


text:
Definition of plaguy in English:
(also plaguey)
adjective[attributive] informal
Troublesome; annoying.
Example sentences
- Today, rare herbs and wild flowers grow on the plaguey pit, so Belfast City Council has ordered the ground left to grow wild in order to see what biological mysteries the cemetery contains.
- Being awake when I should have been asleep was very plaguy.
For editors and proofreaders
Syllabification: pla·guy
Definition of plaguy in:
What do you find interesting about this word or phrase?
Comments that don't adhere to our Community Guidelines may be moderated or removed.
= trending