Pathogen- and Host-Directed Antileishmanial Effects Mediated by Polyhexanide (PHMB)

BACKGROUND:Cutaneous leishmaniasis (CL) is a neglected tropical disease caused by protozoan parasites of the genus Leishmania. CL causes enormous suffering in many countries worldwide. There is no licensed vaccine against CL, and the chemotherapy options show limited efficacy and high toxicity. Localization of the parasites inside host cells is a barrier to most standard chemo- and immune-based interventions. Hence, novel drugs, which are safe, effective and readily accessible to third-world countries and/or drug delivery technologies for effective CL treatments are desperately needed. METHODOLOGY/PRINCIPAL FINDINGS:Here we evaluated the antileishmanial properties and delivery potential of polyhexamethylene biguanide (PHMB; polyhexanide), a widely used antimicrobial and wound antiseptic, in the Leishmania model. PHMB showed an inherent antileishmanial activity at submicromolar concentrations. Our data revealed that PHMB kills Leishmania major (L. major) via a dual mechanism involving disruption of membrane integrity and selective chromosome condensation and damage. PHMB's DNA binding and host cell entry properties were further exploited to improve the delivery and immunomodulatory activities of unmethylated cytosine-phosphate-guanine oligodeoxynucleotides (CpG ODN). PHMB spontaneously bound CpG ODN, forming stable nanopolyplexes that enhanced uptake of CpG ODN, potentiated antimicrobial killing and reduced host cell toxicity of PHMB. CONCLUSIONS:Given its low cost and long history of safe topical use, PHMB holds promise as a drug for CL therapy and delivery vehicle for nucleic acid immunomodulators

Host-parasite co-metabolic activation of antitrypanosomal aminomethyl-benzoxaboroles

<div><p>Recent development of benzoxaborole-based chemistry gave rise to a collection of compounds with great potential in targeting diverse infectious diseases, including human African Trypanosomiasis (HAT), a devastating neglected tropical disease. However, further medicinal development is largely restricted by a lack of insight into mechanism of action (MoA) in pathogenic kinetoplastids. We adopted a multidisciplinary approach, combining a high-throughput forward genetic screen with functional group focused chemical biological, structural biology and biochemical analyses, to tackle the complex MoAs of benzoxaboroles in <i>Trypanosoma brucei</i>. We describe an oxidative enzymatic pathway composed of host semicarbazide-sensitive amine oxidase and a trypanosomal aldehyde dehydrogenase TbALDH3. Two sequential reactions through this pathway serve as the key underlying mechanism for activating a series of 4-aminomethylphenoxy-benzoxaboroles as potent trypanocides; the methylamine parental compounds as pro-drugs are transformed first into intermediate aldehyde metabolites, and further into the carboxylate metabolites as effective forms. Moreover, comparative biochemical and crystallographic analyses elucidated the catalytic specificity of TbALDH3 towards the benzaldehyde benzoxaborole metabolites as xenogeneic substrates. Overall, this work proposes a novel drug activation mechanism dependent on both host and parasite metabolism of primary amine containing molecules, which contributes a new perspective to our understanding of the benzoxaborole MoA, and could be further exploited to improve the therapeutic index of antimicrobial compounds.</p></div

Differential impact of LPG-and PG-deficient Leishmania major mutants on the immune response of human dendritic cells

<div><p>Background</p><p><i>Leishmania major</i> infection induces robust interleukin-12 (IL12) production in human dendritic cells (hDC), ultimately resulting in Th1-mediated immunity and clinical resolution. The surface of <i>Leishmania</i> parasites is covered in a dense glycocalyx consisting of primarily lipophosphoglycan (LPG) and other phosphoglycan-containing molecules (PGs), making these glycoconjugates the likely pathogen-associated molecular patterns (PAMPS) responsible for IL12 induction.</p><p>Methodology/Principal Findings</p><p>Here we explored the role of parasite glycoconjugates on the hDC IL12 response by generating <i>L</i>. <i>major</i> Friedlin V1 mutants defective in LPG alone, (FV1 <i>lpg1-</i>), or generally deficient for all PGs, (FV1 <i>lpg2-</i>). Infection with metacyclic, infective stage, <i>L</i>. <i>major</i> or purified LPG induced high levels of <i>IL12B</i> subunit gene transcripts in hDCs, which was abrogated with FV1 <i>lpg1-</i> infections. In contrast, hDC infections with FV1 <i>lpg2-</i> displayed increased <i>IL12B</i> expression, suggesting other PG-related/<i>LPG2</i> dependent molecules may act to dampen the immune response. Global transcriptional profiling comparing WT, FV1 <i>lpg1-</i>, FV1 <i>lpg2-</i> infections revealed that FV1 <i>lpg1-</i> mutants entered hDCs in a silent fashion as indicated by repression of gene expression. Transcription factor binding site analysis suggests that LPG recognition by hDCs induces IL-12 in a signaling cascade resulting in Nuclear Factor κ B (NFκB) and Interferon Regulatory Factor (IRF) mediated transcription.</p><p>Conclusions/Significance</p><p>These data suggest that <i>L</i>. <i>major</i> LPG is a major PAMP recognized by hDC to induce IL12-mediated protective immunity and that there is a complex interplay between PG-baring <i>Leishmania</i> surface glycoconjugates that result in modulation of host cellular IL12.</p></div

Disturbance and management effects on forest soil organic carbon stocks in the Pacific Northwest

Carbon (C)‐informed forest management requires understanding how disturbance and management influence soil organic carbon (SOC) stocks at scales relevant to landowners and forest policy and management professionals. The continued growth of data sets and publications allows powerful synthesis approaches to be applied to such questions at increasingly fine scales. Here, we report results from a synthesis that used meta‐analysis of published studies and two large observational databases to quantify disturbance and management impacts on SOC stocks. We conducted this, the third in a series of ecoregional SOC assessments, for the Pacific Northwest, which comprises ~8% of the land area but ~12% of the U.S. forest sector C sink. At the ecoregional level, our analysis indicated that fundamental patterns of vegetation, climate, and topography are far more important controls on SOC stocks than land use history, disturbance, or management. However, the same patterns suggested that increased warming, drying, wildland fire, and forest regeneration failure pose significant risks to SOC stocks across the region. Detailed meta‐analysis results indicated that wildfires diminished SOC stocks throughout the soil profile, while prescribed fire only influenced surface organic materials and harvesting had no significant overall impact on SOC. Independent observational data corroborated the negative influence of fire on SOC derived from meta‐analysis, suggested that harvest impacts may vary subregionally with climate or vegetation, and revealed that forests with agricultural uses (e.g., grazing) or legacies (e.g., cultivation) had smaller SOC stocks. We also quantified effects of a range of common forest management practices having either positive (organic amendments, nitrogen [N]‐fixing vegetation establishment, inorganic N fertilization) or no overall effects on SOC (other inorganic fertilizers, urea fertilization, competition suppression through herbicides). In order to maximize the management applications of our results, we qualified them with ratings of confidence based on degree of support across approaches. Last, similar to earlier published assessments from other ecoregions, we supplemented our quantitative synthesis results with a literature review to arrive at a concise set of tactics for adapting management operations to site‐specific criteria.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/174845/1/eap2611_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/174845/2/eap2611-sup-0001-Appendix_S1.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/174845/3/eap2611.pd

A família Rubiaceae na Reserva Biológica Guaribas, Paraíba, Brasil: subfamília Rubioideae The family Rubiaceae in the Guaribas Biological Reserve, Paraíba State, Brazil: subfamily Rubioideae

Este trabalho consiste no levantamento dos representantes de Rubiaceae subfamília Rubioideae na Reserva Biológica Guaribas, Estado da Paraíba, Brasil. Foram realizadas coletas intensivas no período de outubro/2000 a outubro/2001, as quais resultaram em 17 espécies e nove gêneros de Rubioideae. Os gêneros com maior número de espécies foram Psychotria L. (seis) e BorreriaG. Mey. (quatro). Coccocypselum P. Browne, Declieuxia Kunth, Diodia L., Mitracarpus Zucc. ex Roem. & Schult., PalicoureaAubl., RichardiaL. e Staelia Cham. & Schltdl. apresentaram uma única espécie cada. Perama hirsuta Aubl., de posição taxonômica incerta na família, também foi tratada neste trabalho. São apresentados chave, descrições, comentários e ilustrações dos táxons.<br>This paper describes the results of a survey of the Rubiaceae subfamily Rubioideae in the Guaribas Biological Reserve, State of Paraíba, Brazil. Intensive collections were made from October/2000 to October/2001, revealing 17 species and nine genera of Rubioideae. The most diverse genera were Psychotria L. (six species) and Borreria G. Mey. (four species). Coccocypselum P. Browne, Declieuxia Kunth, Diodia L., Mitracarpus Zucc. ex Roem. & Schult., Palicourea Aubl., Richardia L. and Staelia Cham. & Schltdl. were each represented by a single species. Perama hirsuta Aubl., of uncertain position within the family, is also treated in this paper. A key, descriptions, notes, and illustrations of the taxa are provided

SIAARTI guidelines for safety in locoregional anaesthesia

SIAARTI guidelines for safety in locoregional anaesthesi