ABSTRACT
Current antileishmanial treatment is hampered by limitations, such as drug toxicity and the risk of treatment failure, which may be related to parasitic drug resistance. Given the urgent need for novel drugs, the Drugs for Neglected Diseases initiative (DNDi) has undertaken a drug discovery program, which has resulted in the identification of aminopyrazoles, a highly promising antileishmanial chemical series. Multiple experiments have been performed to anticipate the propensity for resistance development. Resistance selection was performed by successive exposure of Leishmania infantum promastigotes (in vitro) and intracellular amastigotes (both in vitro and in golden Syrian hamsters). The stability of the resistant phenotypes was assessed after passage in mice and Lutzomyia longipalpis sandflies. Whole-genome sequencing (WGS) was performed to identify mutated genes, copy number variations (CNVs), and somy changes. The potential role of efflux pumps (the MDR and MRP efflux pumps) in the development of resistance was assessed by coincubation of aminopyrazoles with specific efflux pump inhibitors (verapamil, cyclosporine, and probenecid). Repeated drug exposure of amastigotes did not result in the emergence of drug resistance either in vitro or in vivo Selection at the promastigote stage, however, was able to select for parasites with reduced susceptibility (resistance index, 5.8 to 24.5). This phenotype proved to be unstable after in vivo passage in mice and sandflies, suggesting that nonfixed alterations are responsible for the elevated resistance. In line with this, single nucleotide polymorphisms and indels identified by whole-genome sequencing could not be directly linked to the decreased drug susceptibility. Copy number variations were absent, whereas somy changes were detected, which may have accounted for the transient acquisition of resistance. Finally, aminopyrazole activity was not influenced by the MDR and MRP efflux pump inhibitors tested. The selection performed does not suggest the rapid development of resistance against aminopyrazoles in the field. Karyotype changes may confer elevated levels of resistance, but these do not seem to be stable in the vertebrate and invertebrate hosts. MDR/MRP efflux pumps are not likely to significantly impact the activity of the aminopyrazole leads.
Subject(s)
Antiprotozoal Agents , Drug Resistance , Leishmania infantum , Pyrazoles/pharmacology , Animals , Antiprotozoal Agents/pharmacology , Cricetinae , DNA Copy Number Variations , Drug Resistance/genetics , Leishmania infantum/drug effects , Leishmania infantum/genetics , MiceABSTRACT
OBJECTIVES: Former studies demonstrated quick selection of paromomycin resistance for Leishmania infantum and Leishmania donovani accompanied by increased fitness. The present study aimed to interpret these findings in an epidemiological context by comparing infection of WT and experimentally derived paromomycin-resistant strains in the sand fly vector. METHODS: Depending on the Leishmania species, Lutzomyia longipalpis and Phlebotomus perniciosus or Phlebotomus argentipes sand flies were artificially infected with procyclic promastigotes of WT and paromomycin-resistant L. infantum (MHOM/FR/96/LEM3323-cl4) or L. donovani (MHOM/NP/03/BPK275/0-cl18). The infection rate and gut/stomodeal valve colonization were determined to monitor parasite phenotypic behaviour within the vector. The impact of the previously described gain of fitness in the vertebrate host on infectivity for the vector was assessed by feeding L. longipalpis on Syrian golden hamsters heavily infected with either WT or paromomycin-resistant parasites. RESULTS: WT and paromomycin-resistant Leishmania of both species behaved similarly in terms of infection and parasite location within the studied sand fly species. Blood feeding on infected hamsters did not reveal differences in acquisition of WT and paromomycin-resistant parasites, despite the higher organ burdens observed for the paromomycin-resistant strain. Strains remained resistant after passage in the vector. CONCLUSIONS: Although paromomycin-resistant parasites show an increased parasite fitness in vitro and in laboratory rodents, the intrinsic infection potential of paromomycin-resistant parasites remains unaltered in the sand fly. Of importance is the fact that paromomycin-resistant Leishmania are able to complete development in the natural vectors and produce stomodeal infection with metacyclic forms, which clearly suggests their potential to spread and circulate in nature.
Subject(s)
Leishmania donovani , Leishmania infantum , Phlebotomus , Psychodidae , Animals , Cricetinae , Paromomycin/pharmacologyABSTRACT
Leishmaniasis is a neglected parasitic disease for which the current antileishmania therapeutics are hampered by drug toxicity, high cost, need for parenteral administration, increasing treatment failure rates, and emergence of drug resistance. The R&D pipeline had run fairly dry for several years, but fortunately some new drug candidates are now under (pre)clinical development. Identification of novel drugs will nevertheless remain essential to adequately sustain and improve effective disease control in the future. In this review, a package of standard and accessible R&D approaches is discussed with expansion to some alternative strategies focusing on parasite-host and vector-host interactions.
Subject(s)
Antiprotozoal Agents/pharmacology , Drug Discovery , Leishmania/drug effects , Animals , Drug Resistance , Humans , Leishmania/growth & development , Leishmaniasis/drug therapy , Leishmaniasis/parasitologyABSTRACT
Although miltefosine (MIL) has only been approved for the treatment of visceral leishmaniasis (VL) in 2002, its application in monotherapy already led to the development of two confirmed MIL-resistant isolates by 2009. Although liposomal amphotericin B is recommended as first-line treatment in Europe, MIL is still occasionally used in HIV co-infected patients. Since their immune system is incapable of controlling the infection, high parasite burdens and post-treatment relapses are common. Linked to the particular pharmacokinetic profile of MIL, successive treatment of recurrent relapses could in principle facilitate the emergence of drug resistance. This study evaluated the effect of immunosuppression (cyclophosphamide 150â¯mg/kg once weekly) on the development of MIL-resistance in Syrian golden hamsters infected with Leishmania infantum. The hamsters were treated with MIL (20â¯mg/kg orally for 5 days) whenever clinical signs of infection or relapse were observed. The immunosuppression resulted in a significant depletion of CD4+ lymphocytes and MHCII-expressing cells in peripheral blood, and a concomitant increase in tissue parasite burdens and shorter time to relapse, but the strain's susceptibility upon repeated MIL exposure remained unaltered. This study demonstrates that immunosuppression accelerates the occurrence of relapse without expediting MIL resistance development.
Subject(s)
Antiprotozoal Agents/pharmacology , Drug Resistance , Immunocompromised Host , Leishmania donovani/drug effects , Leishmaniasis, Visceral/drug therapy , Phosphorylcholine/analogs & derivatives , Animals , CD4-Positive T-Lymphocytes/drug effects , Cricetinae , Cyclophosphamide/administration & dosage , Female , Immunosuppressive Agents/administration & dosage , Mesocricetus , Mice , Parasitic Sensitivity Tests , Phosphorylcholine/pharmacology , RecurrenceABSTRACT
Streptococcus pneumoniae, the most common cause of bacterial pneumonia, has developed a wide range of virulence factors to evade the immune system of which the polysaccharide capsule is the most important one. Formation of this capsule is dependent on the cps gene locus, but also involves other genes-like galU. The pyrophosphorylase encoded by galU plays a role in the UDP-glucose metabolism of prokaryotes and is required for the biosynthesis of capsular polysaccharides. In this paper, the effect of a galU mutation leading to a dysfunctional UDP-glucose pyrophosphorylase (UDPG:PP) on in vitro biofilm biomass, adherence to lung epithelial cells and macrophage phagocytosis is studied. Last, in vivo virulence using a Galleria mellonella model has been studied. We show that the mutation improves streptococcal adherence to epithelial cells and macrophage phagocytosis in vitro, while there is no definitive correlation on biofilm formation between parent and mutant strains. Moreover, in vivo virulence is attenuated for all mutated strains. Together, these results demonstrate that a galU mutation in S. pneumoniae influences host cell interactions in vitro and in vivo and can strongly influence the outcome of a streptococcal infection. As such, UDPG:PP is worth investigating further as a potential drug target.
Subject(s)
Bacterial Adhesion , Biofilms/growth & development , Mutant Proteins/genetics , Mutation , Phagocytosis , Streptococcus pneumoniae/enzymology , UTP-Glucose-1-Phosphate Uridylyltransferase/genetics , Animals , Bacterial Capsules/metabolism , Cell Line , Disease Models, Animal , Epithelial Cells/microbiology , Humans , Lepidoptera , Macrophages/immunology , Macrophages/microbiology , Mice , Mutant Proteins/metabolism , Pneumococcal Infections/microbiology , Pneumococcal Infections/pathology , Streptococcus pneumoniae/genetics , Streptococcus pneumoniae/immunology , Streptococcus pneumoniae/physiology , UTP-Glucose-1-Phosphate Uridylyltransferase/metabolism , Virulence Factors/genetics , Virulence Factors/metabolismABSTRACT
Primary mouse macrophages are frequently used to provide an in vitro intracellular model to evaluate antileishmanial drug efficacy. The present study compared the phenotypic characteristics of Swiss, BALB/c, and C57BL/6 mouse bone marrow-derived macrophages and peritoneal exudate cells using different stimulation and adherence protocols upon infection with a Leishmania infantum laboratory strain and two clinical isolates. Evaluation parameters were susceptibility to infection, permissiveness to amastigote multiplication, and impact on drug efficacy. Observed variations in infection of peritoneal exudate cells can mostly be linked to changes in the inflammatory cytokine profiles (IL-6, TNF-α, KC/GRO) rather than to differences in initial production of nitric oxide and reactive oxygen species. Optimization of the cell stimulation and adherence conditions resulted in comparable infection indices among peritoneal exudate cells and the various types of bone marrow-derived macrophages. BALB/c-derived bone marrow-derived macrophages were slightly more permissive to intracellular amastigote replication. Evaluation of antileishmanial drug potency in the various cell systems revealed minimal variation for antimonials and paromomycin, and no differences for miltefosine and amphotericin B. The study results allow to conclude that drug evaluation can be performed in all tested primary macrophages as only marginal differences are observed in terms of susceptibility to infection and impact of drug exposure. Combined with some practical considerations, the use of 24-h starch-stimulated, 48-h adhered, Swiss-derived peritoneal exudate cells can be advocated as an efficient, reliable, relatively quick, and cost-effective tool for routine drug susceptibility testing in vitro whenever the use of primary cells is feasible.
Subject(s)
Antiprotozoal Agents/therapeutic use , Leishmania infantum/drug effects , Leishmania infantum/immunology , Leishmaniasis/drug therapy , Leishmaniasis/immunology , Macrophages/immunology , Amphotericin B/therapeutic use , Animals , Antimony/therapeutic use , Cells, Cultured , Drug Resistance/genetics , Female , Interleukin-6/blood , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Nitric Oxide/metabolism , Parasitic Sensitivity Tests , Paromomycin/therapeutic use , Phosphorylcholine/analogs & derivatives , Phosphorylcholine/therapeutic use , Reactive Oxygen Species/metabolism , Tumor Necrosis Factor-alpha/bloodABSTRACT
OBJECTIVES: Three new chemical series (bicyclic nitroimidazoles, aminopyrazoles and oxaboroles) were selected by Drugs for Neglected Diseases initiative as potential new drug leads for leishmaniasis. Pharmacodynamics studies included both in vitro and in vivo efficacy, cross-resistance profiling against the current antileishmanial reference drugs and evaluation of their cidal activity potential. METHODS: Efficacy against the reference laboratory strains of Leishmania infantum (MHOM/MA(BE)/67/ITMAP263) and L. donovani (MHOM/ET/67/L82) was evaluated in vitro on intracellular amastigotes and in vivo in the early curative hamster model. Cidal activity was assessed over a period of 15 days in an in vitro 'time-to-kill' assay. Cross-resistance was assessed in vitro on a panel of L. infantum strains with different degrees of resistance to either antimony, miltefosine or paromomycin. RESULTS: All lead compounds showed potent and selective in vitro activity against the Leishmania strains tested and no cross-resistance could be demonstrated against any of the current antileishmanial drugs. Cidal activity was obtained in vitro for all series within 15 days of exposure with some differences noted between L. donovani and L. infantum. When evaluated in vivo, all lead compounds showed high efficacy and no adverse effects were observed. CONCLUSIONS: The new lead series were shown to have cidal pharmacodynamic activity. The absence of cross-resistance with any of the current antileishmanial drugs opens possibilities for combination treatment to reduce the likelihood of treatment failures and drug resistance.
Subject(s)
Antiprotozoal Agents/administration & dosage , Antiprotozoal Agents/pharmacology , Leishmania donovani/drug effects , Leishmania infantum/drug effects , Leishmaniasis/drug therapy , Animals , Antimony/pharmacokinetics , Antimony/pharmacology , Antiprotozoal Agents/pharmacokinetics , Boron Compounds/administration & dosage , Boron Compounds/pharmacokinetics , Boron Compounds/pharmacology , Cricetinae , Female , Inhibitory Concentration 50 , Leishmaniasis/parasitology , Mice , Nitroimidazoles/administration & dosage , Nitroimidazoles/pharmacokinetics , Nitroimidazoles/pharmacology , Parasitic Sensitivity Tests , Pyrazoles/administration & dosage , Pyrazoles/pharmacokinetics , Pyrazoles/pharmacologyABSTRACT
For decades antimonials were the drugs of choice for the treatment of visceral leishmaniasis (VL), but the recent emergence of resistance has made them redundant as first-line therapy in the endemic VL region in the Indian subcontinent. The application of other drugs has been limited due to adverse effects, perceived high cost, need for parenteral administration and increasing rate of treatment failures. Liposomal amphotericin B (AmB) and miltefosine (MIL) have been positioned as the effective first-line treatments; however, the number of monotherapy MIL-failures has increased after a decade of use. Since no validated molecular resistance markers are yet available, monitoring and surveillance of changes in drug sensitivity and resistance still depends on standard phenotypic in vitro promastigote or amastigote susceptibility assays. Clinical isolates displaying defined MIL- or AmB-resistance are still fairly scarce and fundamental and applied research on resistance mechanisms and dynamics remains largely dependent on laboratory-generated drug resistant strains. This review addresses the various challenges associated with drug susceptibility and -resistance monitoring in VL, with particular emphasis on the choice of strains, susceptibility model selection and standardization of procedures with specific read-out parameters and well-defined threshold criteria. The latter are essential to support surveillance systems and safeguard the limited number of currently available antileishmanial drugs.
Subject(s)
Antiprotozoal Agents/adverse effects , Drug Resistance, Multiple , Leishmania donovani/drug effects , Leishmaniasis, Visceral/drug therapy , Parasitic Sensitivity Tests/standards , Amphotericin B/administration & dosage , Amphotericin B/adverse effects , Amphotericin B/therapeutic use , Animals , Antiprotozoal Agents/pharmacology , Antiprotozoal Agents/therapeutic use , Humans , Leishmaniasis, Visceral/epidemiology , Leishmaniasis, Visceral/parasitology , Meglumine Antimoniate/adverse effects , Meglumine Antimoniate/therapeutic use , Parasitic Sensitivity Tests/methods , Phosphorylcholine/analogs & derivatives , Phosphorylcholine/pharmacology , Phosphorylcholine/therapeutic use , Psychodidae/parasitology , RecurrenceABSTRACT
Respiratory syncytial virus (RSV) infections remain a major cause of respiratory disease and hospitalizations among infants. Infection recurs frequently and establishes a weak and short-lived immunity. To date, RSV immunoprophylaxis and vaccine research is mainly focused on the RSV fusion (F) protein, but a vaccine remains elusive. The RSV F protein is a highly conserved surface glycoprotein and is the main target of neutralizing antibodies induced by natural infection. Here, we analyzed an internalization process of antigen-antibody complexes after binding of RSV-specific antibodies to RSV antigens expressed on the surface of infected cells. The RSV F protein and attachment (G) protein were found to be internalized in both infected and transfected cells after the addition of either RSV-specific polyclonal antibodies (PAbs) or RSV glycoprotein-specific monoclonal antibodies (MAbs), as determined by indirect immunofluorescence staining and flow-cytometric analysis. Internalization experiments with different cell lines, well-differentiated primary bronchial epithelial cells (WD-PBECs), and RSV isolates suggest that antibody internalization can be considered a general feature of RSV. More specifically for RSV F, the mechanism of internalization was shown to be clathrin dependent. All RSV F-targeted MAbs tested, regardless of their epitopes, induced internalization of RSV F. No differences could be observed between the different MAbs, indicating that RSV F internalization was epitope independent. Since this process can be either antiviral, by affecting virus assembly and production, or beneficial for the virus, by limiting the efficacy of antibodies and effector mechanism, further research is required to determine the extent to which this occurs in vivo and how this might impact RSV replication.IMPORTANCE Current research into the development of new immunoprophylaxis and vaccines is mainly focused on the RSV F protein since, among others, RSV F-specific antibodies are able to protect infants from severe disease, if administered prophylactically. However, antibody responses established after natural RSV infections are poorly protective against reinfection, and high levels of antibodies do not always correlate with protection. Therefore, RSV might be capable of interfering, at least partially, with antibody-induced neutralization. In this study, a process through which surface-expressed RSV F proteins are internalized after interaction with RSV-specific antibodies is described. One the one hand, this antigen-antibody complex internalization could result in an antiviral effect, since it may interfere with virus particle formation and virus production. On the other hand, this mechanism may also reduce the efficacy of antibody-mediated effector mechanisms toward infected cells.
Subject(s)
Antibodies, Viral/metabolism , Endocytosis , Respiratory Syncytial Virus, Human/immunology , Viral Fusion Proteins/metabolism , Antigen-Antibody Complex/metabolism , Cells, Cultured , Flow Cytometry , Fluorescent Antibody Technique, Indirect , HumansABSTRACT
This study reports the evaluation of the technical and clinical validation of the O-DiaBorburg kit (DIA), Borrelia burgdorferi PCR kit, ISEX (GENE), and Borrelia burgdorferi sensu lato Real-TM (SAC) for the diagnosis of neuroborreliosis in cerebrospinal fluid based on both Borrelia DNA and CSF samples from patients with clinical suspicion of neuroborreliosis. This validation study was done by analysing the kits on the Rotorgene Q (RGQ), CFX96, and LightCycler480 (LC480). For all kits, the linear range was larger on RGQ than on CFX96 and LC480. A good reproducibility was obtained for all assays on all instruments. Storage at -20 °C resulted in a decreased reproducibility for SAC. Results of the limit of detection (LOD95) experiments indicated a better sensitivity than described in the kit insert for all kits on all PCR platforms. No cross-reactivity was found for genetically related organisms nor for other pathogens which may be present in CSF. All species of the Borrelia burgdorferi sensu lato complex were detected with the GENE and SAC kits. The DIA kit failed to detect B. lusitaniae. The results seemed to indicate a better overall performance for the GENE kit on RGQ. However, its diagnostic value could not be confirmed in the clinical validation study, wherein none of the 103 CSF samples from clinical neuroborreliosis cases showed a positive real-time PCR result with the GENE kit analysed on RGQ.
Subject(s)
Borrelia burgdorferi Group/isolation & purification , Cerebrospinal Fluid/microbiology , Lyme Neuroborreliosis/diagnosis , Molecular Diagnostic Techniques/methods , Real-Time Polymerase Chain Reaction/methods , Adolescent , Adult , Aged , Aged, 80 and over , Borrelia burgdorferi Group/genetics , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Reproducibility of Results , Retrospective Studies , Sensitivity and Specificity , Young AdultABSTRACT
BACKGROUND: A short, reliable and valid tool to measure snack and beverage consumption in adolescents, taking into account the correct definitions, would benefit both epidemiological and intervention research. The present study aimed to develop a short quantitative beverage and snack food frequency questionnaire (FFQ) and to assess the reliability and validity of this FFQ against three 24-h recalls. METHODS: Reliability was assessed by comparing estimates of the FFQ administered 14 days apart (FFQ1 and FFQ2) in a convenience sample of 179 adolescents [60.3% male; mean (SD) 14.7 (0.9) years]. Validity was assessed by comparing FFQ1 with three telephone-administered 24-h recalls in a convenience sample of 99 adolescents [52.5% male, mean (SD) 14.8 (0.9) years]. Reliability and validity were assessed using Bland-Altman plots, classification agreements and correlation coefficients for the amount and frequency of consumption of unhealthy snacks, healthy snacks, unhealthy beverages, healthy beverages, and for the healthy snack and beverage ratios. RESULTS: Small mean differences (FFQ1 versus FFQ2) were observed for reliability, ranking ability ranged from fair to substantial, and Spearman coefficients fell within normal ranges. For the validity, mean differences (FFQ1 versus recalls) were small for beverage intake but large for snack intake, except for the healthy snack ratio. Ranking ability ranged from slightly to moderate, and Spearman coefficients fell within normal ranges. CONCLUSIONS: Reliability and validity of the FFQ for all outcomes were found to be acceptable at a group level for epidemiological purposes, whereas for intervention purposes only the healthy snack and beverage ratios were found to be acceptable at a group level.
Subject(s)
Beverages , Diet Records , Snacks , Surveys and Questionnaires , Adolescent , Cross-Sectional Studies , Evaluation Studies as Topic , Female , Humans , Male , Mental Recall , Reproducibility of ResultsABSTRACT
OBJECTIVES: Despite a continued search for novel antileishmanial drugs, treatment options remain restricted to a few standard drugs, e.g. antimonials, miltefosine, amphotericin B and paromomycin. Although these drugs have now been used for several decades, their mechanism of action still remains partly hypothetical and their dynamics of cidal action and time-to-kill are still poorly documented. METHODS: An in vitro time-to-kill assay on intracellular amastigotes of the laboratory reference strains Leishmania donovani (MHOM/ET/67/L82) and Leishmania infantum [MHOM/MA(BE)/67/ITMAP263] evaluated the cidal action dynamics of the listed reference drugs at three different concentrations: at IC50, 2â×âIC50 and the near cytotoxic dose level (CC90: determined on MRC-5 cells). This assay focused on identifying the minimal exposure time needed to completely eliminate viable intracellular amastigotes, using the standard microscopic Giemsa assay and the promastigote back-transformation assay. RESULTS: While 100% reduction was microscopically apparent for most drugs, the promastigote back-transformation assay clearly demonstrated a concentration- and time-dependent cidal mechanism. The time-to-kill at 2â×âIC50 was ≥240 h for pentavalent antimony (77 µg eq./mL), 96 h for trivalent antimony (44 µg eq./mL), 168 to >240 h for miltefosine (10 µM), 168 h for paromomycin (100 µM) and >240 h for amphotericin B (2 µM). No differences were noted between both Leishmania species. CONCLUSIONS: Evaluation of the concentration- and time-dependent cidal activity using the promastigote back-transformation assay revealed striking differences in efficacy of the different antileishmanial reference drugs. This assay should allow in-depth pharmacodynamic evaluation of novel drug leads in comparison with the existing antileishmanial drug repertoire.
Subject(s)
Antiprotozoal Agents/pharmacology , Leishmania donovani/drug effects , Leishmania donovani/physiology , Leishmania infantum/drug effects , Leishmania infantum/physiology , Cell Survival/drug effects , Inhibitory Concentration 50 , Parasitic Sensitivity Tests , Time FactorsABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: The Nkundo people (Nkundo area of Bolongo, Mai-Ndombe district, Bandundu Province, DR Congo) use various plant parts of the tree Greenwayodendron suaveolens (Engl. & Diels) Verdc. (syn. Polyalthia suaveolens Engl. & Diels) (Annonaceae) against malaria, but its antiprotozoal constituents are not known. MATERIALS AND METHODS: The crude 80% ethanol extract from the fruits, leaves, root bark and stem bark and 16 fractions were assessed in vitro for their antiprotozoal activity against Trypanosoma brucei brucei, T. cruzi, Leishmania infantum and the chloroquine and pyrimethamine-resistant K1 strain of Plasmodium falciparum (Pf-K1). Their cytotoxic effects were evaluated against MRC-5 cells. Active constituents were isolated by chromatographic means, identified using spectroscopic methods, and evaluated in the same assays. RESULTS: The root bark extract showed the highest activity against P. falciparum K1 (IC50 0.26µg/mL) along with the stem bark alkaloid fraction (IC50 0.27µg/mL). The root bark alkaloid fraction had a pronounced activity against all selected protozoa with IC50 values <1µg/mL. The 90% methanol fractions of the different plant parts showed a pronounced activity against P. falciparum K1, with IC50 values ranging between 0.36µg/mL and 0.69µg/mL. Four constituents were isolated: the triterpenes polycarpol, and dihydropolycarpol, the latter one being reported for the first time from nature, and the alkaloids polyalthenol and N-acetyl-polyveoline. They were active to a various degree against one or more protozoa, mostly accompanied by cytotoxicity. The highest selectivity was observed for N-acetyl-polyveoline against P. falciparum K1 (IC50 2.8µM, selectivity index 10.9). CONCLUSIONS: These results may explain at least in part the traditional use of this plant species against parasitic diseases such as malaria in DR Congo.
Subject(s)
Antimalarials/pharmacology , Phytochemicals/pharmacology , Plant Extracts/pharmacology , Plasmodium falciparum/drug effects , Polyalthia/chemistry , Trypanocidal Agents/pharmacology , Antimalarials/isolation & purification , Antimalarials/toxicity , Cell Line , Cell Survival/drug effects , Dose-Response Relationship, Drug , Fruit/chemistry , Humans , Inhibitory Concentration 50 , Leishmania infantum/drug effects , Leishmania infantum/growth & development , Parasitic Sensitivity Tests , Phytochemicals/isolation & purification , Phytochemicals/toxicity , Phytotherapy , Plant Bark/chemistry , Plant Extracts/isolation & purification , Plant Extracts/toxicity , Plant Leaves/chemistry , Plant Roots/chemistry , Plants, Medicinal , Plasmodium falciparum/growth & development , Trypanocidal Agents/isolation & purification , Trypanocidal Agents/toxicity , Trypanosoma brucei brucei/drug effects , Trypanosoma brucei brucei/growth & developmentABSTRACT
In this work we present the methodology for the development of the EMBalance diagnostic Decision Support System (DSS) for balance disorders. Medical data from patients with balance disorders have been analysed using data mining techniques for the development of the diagnostic DSS. The proposed methodology uses various data, ranging from demographic characteristics to clinical examination, auditory and vestibular tests, in order to provide an accurate diagnosis. The system aims to provide decision support for general practitioners (GPs) and experts in the diagnosis of balance disorders as well as to provide recommendations for the appropriate information and data to be requested at each step of the diagnostic process. Detailed results are provided for the diagnosis of 12 balance disorders, both for GPs and experts. Overall, the reported accuracy ranges from 59.3 to 89.8% for GPs and from 74.3 to 92.1% for experts.
Subject(s)
Data Mining/methods , Decision Support Systems, Clinical , Decision Support Techniques , Vestibule, Labyrinth/physiology , Algorithms , Decision Trees , Humans , Postural Balance/physiology , Vertigo/diagnosisABSTRACT
BACKGROUND: As the snacking pattern of European adolescents is of great concern, effective interventions are necessary. Till now health promotion efforts in children and adolescents have had only limited success in changing adolescents' eating patterns and anthropometrics. Therefore, the present study proposes an innovative approach to influence dietary behaviors in youth based on new insights on effective behavior change strategies and attractive intervention channels to engage adolescents. This article describes the rationale, the development, and evaluation design of the 'Snack Track School' app. The aim of the app is to improve the snacking patterns of Flemish 14- to 16-year olds. METHODS: The development of the app was informed by the systematic, stepwise, iterative, and collaborative principles of the Intervention Mapping protocol. A four week mHealth intervention was developed based on the dual-system model with behavioral change strategies targeting both the reflective (i.e., active learning, advance organizers, mere exposure, goal-setting, monitoring, and feedback) and automatic processes (i.e., rewards and positive reinforcement). This intervention will be evaluated via a controlled pre-post design in Flemish schools among 1400 adolescents. DISCUSSION: When this intervention including strategies focused on both the reflective and automatic pathway proves to be effective, it will offer a new scientifically-based vision, guidelines and practical tools for public health and health promotion (i.e., incorporation of learning theories in intervention programs). TRIAL REGISTRATION: NCT02622165 registrated November 15, 2015 on clinicaltrials.gov.
Subject(s)
Behavior Therapy , Diet , Feeding Behavior , Health Promotion/methods , Mobile Applications , Reward , Snacks , Adolescent , Belgium , Feedback , Female , Goals , Humans , Learning , Male , Research Design , Schools , Self-Control , Surveys and Questionnaires , Telemedicine , Video GamesABSTRACT
UNLABELLED: Aim/Purpose of the Study: Activation of the renin-angiotensin system leading to increased angiotensin-(1-7) (Ang-(1-7)) and decreased angiotensin 2 (Ang 2) levels may be a new therapeutic approach to reduce acute lung injury. Prolylcarboxypeptidase (PRCP) and prolyloligopeptidase (PREP) are capable of hydrolyzing Ang 2 into Ang-(1-7). However, their relation with circulating Ang 2 levels after lung ischemia-reperfusion injury (LIRI) has never been explored. This study determines whether the activity and expression of PRCP and PREP in plasma and lung tissue is related to circulating Ang 2 levels in a murine model of LIRI. MATERIALS AND METHODS: LIRI in Swiss mice (6 animals per group) was induced by temporary left lung hilar clamping (1 h) followed by 0, 1 or 24 h of reperfusion. Animals in the sham group received thoracotomy only. PRCP activity was measured via RP-HPLC, PREP activity using a fluorogenic substrate and plasma Ang 2 levels via ELISA. Western blotting was used to determine the PRCP and PREP protein expression profiles in left lung tissue. RESULTS: Plasma Ang 2 levels significantly rise after lung ischemia and remain increased after 1 h and 24 h of reperfusion compared to the sham group. While a significant decrease in plasma PREP activity was found after 24 h of reperfusion, a transient increase in plasma PRCP activity was observed after ischemia. However, no correlation with plasma Ang 2 levels could be demonstrated. The activity profiles of PRCP and PREP and the protein expression of PRCP in the lung tissues remained unchanged after LIRI. CONCLUSIONS: LIRI causes a dysregulation of circulating Ang 2 levels and plasma PREP activity, although no direct link between both phenomena could be shown. The activity profile of pulmonary PRCP and PREP was not significantly changed after LIRI, which implies a minor role for local PRCP and PREP in the ischemic lung itself.
Subject(s)
Angiotensin II/blood , Carboxypeptidases/blood , Lung Injury/metabolism , Renin-Angiotensin System , Reperfusion Injury/metabolism , Serine Endopeptidases/blood , Animals , Disease Models, Animal , Female , Lung/enzymology , Lung Injury/physiopathology , Mice , Prolyl Oligopeptidases , Reperfusion Injury/physiopathologyABSTRACT
The Interuniversity Attraction Pole (IAP) 'PLANET TOPERS' (Planets: Tracing the Transfer, Origin, Preservation, and Evolution of their Reservoirs) addresses the fundamental understanding of the thermal and compositional evolution of the different reservoirs of planetary bodies (core, mantle, crust, atmosphere, hydrosphere, cryosphere, and space) considering interactions and feedback mechanisms. Here we present the first results after 2 years of project work.
Subject(s)
Evolution, Planetary , Extraterrestrial Environment , Planets , ExobiologyABSTRACT
OBJECTIVES: Although miltefosine and paromomycin were only recently introduced to treat visceral leishmaniasis, increasing numbers of miltefosine treatment failures and occasional primary resistance to both drugs have been reported. Understanding alterations in parasite behaviour linked to drug resistance is essential to assess the propensity for emergence and spread of resistant strains, particularly since a positive effect on fitness has been reported for antimony-resistant parasites. This laboratory study compared the fitness of a drug-susceptible parent WT clinical Leishmania infantum isolate (MHOM/FR/96/LEM3323) and derived miltefosine and paromomycin drug-resistant lines that were experimentally selected at the intracellular amastigote level. METHODS: Parasite fitness of WT, paromomycin-resistant and miltefosine-resistant strains, in vitro and in vivo parasite growth, metacyclogenesis, infectivity and macrophage stress responses were comparatively evaluated. RESULTS: No significant differences in promastigote fitness were noted between the WT and paromomycin-resistant strain, while clear benefits could be demonstrated for paromomycin-resistant amastigotes in terms of enhanced in vitro and in vivo growth potential and intracellular stress response. The miltefosine-resistant phenotype showed incomplete promastigote metacyclogenesis, decreased intracellular growth and weakened stress response, revealing a reduced fitness compared with WT parent parasites. CONCLUSIONS: The rapid selection and fitness advantages of paromomycin-resistant amastigotes endorse the current use of paromomycin in combination therapy. Although a reduced fitness of miltefosine-resistant strains may explain the difficulty of miltefosine resistance selection in vitro, the growing number of miltefosine treatment failures in the field still requires further exploratory research.
Subject(s)
Antiprotozoal Agents/pharmacology , Drug Resistance , Leishmania infantum/drug effects , Paromomycin/pharmacology , Phosphorylcholine/analogs & derivatives , Animals , Disease Models, Animal , Female , Humans , Leishmania infantum/pathogenicity , Leishmania infantum/physiology , Leishmaniasis, Visceral/parasitology , Leishmaniasis, Visceral/pathology , Mice, Inbred BALB C , Phosphorylcholine/pharmacology , Selection, Genetic , VirulenceABSTRACT
Leishmania donovani is a protozoan parasite causing the neglected tropical disease visceral leishmaniasis. One difficulty to study the immunopathology upon L. donovani infection is the limited adaptability of the strains to experimental mammalian hosts. Our knowledge about L. donovani infections relies on a restricted number of East African strains (LV9, 1S). Isolated from patients in the 1960s, these strains were described extensively in mice and Syrian hamsters and have consequently become 'reference' laboratory strains. L. donovani strains from the Indian continent display distinct clinical features compared to East African strains. Some reports describing the in vivo immunopathology of strains from the Indian continent exist. This study comprises a comprehensive immunopathological characterization upon infection with two additional strains, the Ethiopian L. donovani L82 strain and the Nepalese L. donovani BPK282 strain in both Syrian hamsters and C57BL/6 mice. Parameters that include parasitaemia levels, weight loss, hepatosplenomegaly and alterations in cellular composition of the spleen and liver, showed that the L82 strain generated an overall more virulent infection compared to the BPK282 strain. Altogether, both L. donovani strains are suitable and interesting for subsequent in vivo investigation of visceral leishmaniasis in the Syrian hamster and the C57BL/6 mouse model.
Subject(s)
Leishmania donovani/classification , Leishmaniasis, Visceral/parasitology , Animals , Cricetinae , DNA, Kinetoplast/genetics , DNA, Protozoan/genetics , Humans , Leishmania donovani/genetics , Leishmania donovani/pathogenicity , Leishmaniasis, Visceral/pathology , Liver/parasitology , Liver/pathology , Mesocricetus , Mice , Mice, Inbred C57BL , Spleen/parasitology , Spleen/pathology , VirulenceABSTRACT
Tuberculosis has remained a challenge for medicinal chemists worldwide. In the framework of a collaborative program to identify and evaluate novel antitubercular candidate compounds, the biological properties of benzo[g]isoquinoline-5,10-diones have been found to be very promising. In this paper we have further expanded the library by incorporation of an amidinium moiety into the benzo[g]isoquinoline-5,10-dione scaffold. The presence of this functional group also increased the solubility of the quinones in polar solvents. To this purpose N(2)-arylbenzo[g]isoquinoline-5,10-dione-3-iminium bromides were synthesized in a straightforward way by means of a reaction of anilines with 2-(bromomethyl)-3-(cyanomethyl)-1,4-dimethoxynaphthalene. Following the biological evaluation, N(2)-(4-chlorophenyl)-5,10-dioxobenzo[g]isoquinoline-3(2H)-iminium bromide (MIC = 1.16 µM, CC50 = 28.51 µM, SI = 24.58) was selected as the most promising representative. Apart from the nano-molar anti-mycobacterial activity, the compound was able to target intracellular residing Mycobacterium tuberculosis and the susceptibility of a multi-drug-resistant strain towards the compound was confirmed.