A Cell-surface Phylome for African Trypanosomes

The cell surface of Trypanosoma brucei, like many protistan blood parasites, is crucial for mediating host-parasite interactions and is instrumental to the initiation, maintenance and severity of infection. Previous comparisons with the related trypanosomatid parasites T. cruzi and Leishmania major suggest that the cell-surface proteome of T. brucei is largely taxon-specific. Here we compare genes predicted to encode cell surface proteins of T. brucei with those from two related African trypanosomes, T. congolense and T. vivax. We created a cell surface phylome (CSP) by estimating phylogenies for 79 gene families with putative surface functions to understand the more recent evolution of African trypanosome surface architecture. Our findings demonstrate that the transferrin receptor genes essential for bloodstream survival in T. brucei are conserved in T. congolense but absent from T. vivax and include an expanded gene family of insect stage-specific surface glycoproteins that includes many currently uncharacterized genes. We also identify species-specific features and innovations and confirm that these include most expression site-associated genes (ESAGs) in T. brucei, which are absent from T. congolense and T. vivax. The CSP presents the first global picture of the origins and dynamics of cell surface architecture in African trypanosomes, representing the principal differences in genomic repertoire between African trypanosome species and provides a basis from which to explore the developmental and pathological differences in surface architectures. All data can be accessed at: http://www.genedb.org/Page/trypanosoma_surface_phylome.     

Dual-stage triterpenoids from an African medicinal plant targeting the malaria parasite

Sixteen triterpenoids (1–16), previously isolated from the aerial parts of the African medicinal plant Momordica balsamina or obtained by derivatization, were evaluated for their activity against liver stages of Plasmodium berghei, measuring the luminescence intensity in Huh-7 cells infected with a firefly luciferase-expressing P. berghei line, PbGFP-Luc_con. Toxicity of compounds (1–16) was assessed on the same cell line through the fluorescence measurement of cell confluency. The highest activity was displayed by a derivative bearing two acetyl residues, karavoate B (7), which led to a dose-dependent decrease in the P. berghei infection rate, exhibiting a very significant activity at the lowest concentration employed (1 μM) and no toxicity towards the Huh-7 cells. It is noteworthy that, in previous studies, this compound was found to be a strong inhibitor of blood-stages of Plasmodium falciparum, thus displaying a dual-stage antimalarial activity.     

The genome of Mycobacterium africanum West African 2 reveals a lineage-specific locus and genome erosion common to the M. tuberculosis complex

Background

M. africanum West African 2 constitutes an ancient lineage of the M. tuberculosis complex that commonly causes human tuberculosis in West Africa and has an attenuated phenotype relative to M. tuberculosis.

Methodology/Principal Findings

In search of candidate genes underlying these differences, the genome of M. africanum West African 2 was sequenced using classical capillary sequencing techniques. Our findings reveal a unique sequence, RD900, that was independently lost during the evolution of two important lineages within the complex: the “modern” M. tuberculosis group and the lineage leading to M. bovis. Closely related to M. bovis and other animal strains within the M. tuberculosis complex, M. africanum West African 2 shares an abundance of pseudogenes with M. bovis but also with M. africanum West African clade 1. Comparison with other strains of the M. tuberculosis complex revealed pseudogenes events in all the known lineages pointing toward ongoing genome erosion likely due to increased genetic drift and relaxed selection linked to serial transmission-bottlenecks and an intracellular lifestyle.

Conclusions/Significance

The genomic differences identified between M. africanum West African 2 and the other strains of the Mycobacterium tuberculosis complex may explain its attenuated phenotype, and pave the way for targeted experiments to elucidate the phenotypic characteristic of M. africanum. Moreover, availability of the whole genome data allows for verification of conservation of targets used for the next generation of diagnostics and vaccines, in order to ensure similar efficacy in West Africa.     

Fruit nutritional composition and non-nutritive traits of indigenous South African tree species

Frugivorous animals play a major role in dispersing tropical, and to a lesser extent, temperate tree species. In order to attract potential seed dispersers, plants generally offer a reward of fleshy fruit pulp. Criteria for fruit choice by avian frugivores are influenced by a number of non-nutritive (e.g. fruit size and colour) factors; and nutritional composition of the fruit. There is a paucity of nutritional composition and other fruit trait data of indigenous South African fruit. This information is necessary in order to determine which frugivores are likely to ingest which fruits and consequently act as potential seed dispersal agents. This information would provide us with an understanding of the inter-relationships between indigenous fruit and frugivores in South Africa. Consequently nutritional composition was investigated in various indigenous fruit species that avian frugivores feed on. Fruits were collected from 38 indigenous tree species found in KwaZulu-Natal Afromontane and coastal forests. Pulp was freeze-dried to constant mass and then analysed for sugar, lipid and protein content; and for water content determination. Fruit width in this study ranged from 4 mm (Searsia rehmanniana and Trema orientalis) to 40 mm (Annona senegalensis, Ficus sur and Xylotheca kraussiana). Of the fruits examined in this study 29% were black and 43% were red when ripe. Most (84%) fruit species analysed for sugar content were hexose dominant with 50% being fructose and 34% being glucose dominant. Only 16% of the fruit species analysed were sucrose dominant. Fruits in this study were generally observed to be high (mean: 68.1 ± 3.3%; n = 30) in water content; and low in protein and lipid content respectively (mean: 8.2 ± 0.5%; 9.3 ± 2.2%; n = 30) indicating that these fruit species could be considered as nutrient-dilute. Future studies need to determine the nutritional composition of the remaining indigenous South African fruit in order to develop a comprehensive database as well as examining non-nutritive factors.     

Persistent Association of Mycobacterium ulcerans with West African Predaceous Insects of the Family Belostomatidae

A number of studies have suggested that Mycobacterium ulcerans, the etiological agent of Buruli ulcer, may be transmitted to humans by insect bites. M. ulcerans has been isolated from a predaceous aquatic insect, and PCR detection of M. ulcerans DNA in aquatic environments suggests that the organism is widely distributed within many invertebrate taxa and functional feeding groups. Thus, M. ulcerans may be concentrated through different trophic links. However, the specific environmental niche of M. ulcerans and route of transmission to humans remain a mystery. In this study, a biologically relevant infection model in which M. ulcerans-infected mosquito larvae were fed to a species of predaceous hemiptera (African Belostomatidae) was used to demonstrate the persistent colonization of M. ulcerans and subsequent transmission of bacteria to naïve prey. The association of M. ulcerans with specific anatomical compartments showed that M. ulcerans accumulates preferentially on the exoskeleton. In contrast, few organisms were found in dissected guts or salivary glands. No difference was found between the ability of wild-type M. ulcerans and an M. ulcerans isogenic mycolactone-negative mutant to colonize belostomatids. These data show that African belostomatids can successfully be colonized by M. ulcerans and support the trophic transfer of M. ulcerans within the environment.     

Henipavirus RNA in African Bats

Background

Henipaviruses (Hendra and Nipah virus) are highly pathogenic members of the family Paramyxoviridae. Fruit-eating bats of the Pteropus genus have been suggested as their natural reservoir. Human Henipavirus infections have been reported in a region extending from Australia via Malaysia into Bangladesh, compatible with the geographic range of Pteropus. These bats do not occur in continental Africa, but a whole range of other fruit bats is encountered. One of the most abundant is Eidolon helvum, the African Straw-coloured fruit bat.

Methodology/Principal Findings

Feces from E. helvum roosting in an urban setting in Kumasi/Ghana were tested for Henipavirus RNA. Sequences of three novel viruses in phylogenetic relationship to known Henipaviruses were detected. Virus RNA concentrations in feces were low.

Conclusions/Significance

The finding of novel putative Henipaviruses outside Australia and Asia contributes a significant extension of the region of potential endemicity of one of the most pathogenic virus genera known in humans.     

Analysis of N-glycan profile of Arabidopsis alg3 cell culture

N-Glycosylation is essential for protein stability, activity and characteristics, and is often needed to deliver pharmaceutical glycoproteins to target cells. A paucimannosidic structure, Man₃GlcNAc₂ (M3), has been reported to enable cellular uptake of glycoproteins through the mannose receptor (MR) in humans, and such uptake has been exploited for the treatment of certain diseases. However, M3 is generally produced at a very low level in plants. In this study, a cell culture was established from an Arabidopsis alg3 mutant plant lacking asparagine-linked glycosylation 3 (ALG3) enzyme activity. Arabidopsis alg3 cell culture produced glycoproteins with predominantly M3 and GlcNAc-terminal structures, while the amount of plant-specific N-glycans was very low. Pharmaceutical glycoproteins with these characteristics would be valuable for cellular delivery through the MR, and safe for human therapy.     

Phylogenetic position of African and Malagasy Pimpinella species and related genera (Apiaceae, Pimpinelleae)

The phylogenetic position of the African and Malagasy species of Pimpinella is assessed using nrDNA ITS sequence data and a representative sampling of the genus, including 16 species from Africa and Madagascar and 26 species from Eurasia. The results of maximum parsimony and Bayesian analyses of these data show that the African and Malagasy species ally with their Eurasian counterparts in Pimpinelleae. The genus Pimpinella is rendered paraphyletic by the inclusion of African Cryptotaenia and the small African and Malagasy endemic genera Frommia and Phellolophium. Within a paraphyletic Pimpinella, three major clades are recovered, with the African species occupying two of these clades. The current sectional classification of the genus, based predominantly on fruit vestiture, is largely artificial. Chromosome base number, however, was found to be consistent with the groupings recovered in the molecular analyses. Those African and Malagasy Pimpinella species with a chromosome base number of x = 11 and largely glabrous petals and fruits, form the earliest diverging clade together with Frommia, which also has a base count of n = 11 and glabrous petals and fruits. The remaining African species ally with several Eurasian species of Pimpinella and share a chromosome base number of x = 9 and usually hairy petals and fruits.     

Pancreatic Carcinoma in an African Clawed Frog (Xenopus laevis)

This report describes the histologic features of a pancreatic carcinoma in an adult female African clawed frog (Xenopus laevis). The animal was found to be in poor body condition and subsequently euthanized for a complete necropsy. Histologically, the pancreas was effaced by packets of polyhedral cells consistent with a pancreatic islet cell carcinoma. Metastatic disease was not identified. Pancreatic tumors are uncommon in amphibians, and this report is the first to describe a pancreatic carcinoma in an African clawed frog.The African clawed frog (Xenopus laevis) is one of the world''s most commonly used frog species in developmental biology, cell biology, toxicology, and neuroscience. Neoplastic diseases in Xenopus have rarely been reported in the literature but include hepatomas, teratomas, renal carcinoma, fibroma, fibrosarcoma, nephroblastoma, ovarian dysgerminoma, melanophoromas and lymphoma.^{1,2,5,6,10,14} Although hyperplasia of pancreatic islets in Xenopus has been described, pancreatic neoplasms in these frogs have not previously been reported.¹⁵ Pancreatic carcinomas have been reported to occur in hybrid species of pond frogs of the Pelophylax (previously Rana) genus.⁹ This report describes the first documented occurrence of a pancreatic carcinoma in X. laevis.     

Functional Rectification of the Newly Described African Henipavirus Fusion Glycoprotein (Gh-M74a)

Recent evidence identified multiple Henipavirus species in Africa distinct from those in Southeast Asia and Australia. The reported fusion glycoprotein (F) sequence of the African Gh-M74a strain (GhV-F) is likely incorrect: a single base pair deletion near the N terminus results in multiple aberrancies. Rectifying this by adding single nucleotide insertions results in a GhV-F that now possesses a signal peptide, is efficiently cell surface expressed, exhibits syncytium formation when coexpressed with GhV-G protein, and mediates pseudotyped viral particle entry.     

African Origin and Europe-Mediated Global Dispersal of The Cyanobacterium Microcystis aeruginosa

Microcystis aeruginosa is a bloom-forming cyanobacteria, which currently has a cosmopolitan distribution. Since M. aeruginosa can produce toxic compounds across all continents that it inhabits, it is of major public health relevance to assess its origin and dispersal. Thus, we conducted a worldwide study using 29 isolates representative of all the main continents, and used a concatenated genetic system for phylogenetic analyses consisting of four genetic markers (spanning ca. 3,485 bp). Our results support an early origin of M. aeruginosa in the African continent, with a subsequent dispersal to establish a second genetic pool in the European continent, from where M. aeruginosa then colonized the remaining continental regions. Our findings indicate that the European population has a cosmopolitan distribution, and is genetically closer to populations from Africa and North America. Our study also highlights the utility of using a concatenated dataset for phylogenetic inferences in cyanobacteria.     

African ancestry is associated with asthma risk in African Americans

Background

Asthma is a common complex condition with clear racial and ethnic differences in both prevalence and severity. Asthma consultation rates, mortality, and severe symptoms are greatly increased in African descent populations of developed countries. African ancestry has been associated with asthma, total serum IgE and lower pulmonary function in African-admixed populations. To replicate previous findings, here we aimed to examine whether African ancestry was associated with asthma susceptibility in African Americans. In addition, we examined for the first time whether African ancestry was associated with asthma exacerbations.

Methodology/Principal Findings

After filtering for self-reported ancestry and genotype data quality, samples from 1,117 self-reported African-American individuals from New York and Baltimore (394 cases, 481 controls), and Chicago (321 cases followed for asthma exacerbations) were analyzed. Genetic ancestry was estimated based on ancestry informative markers (AIMs) selected for being highly divergent among European and West African populations (95 AIMs for New York and Baltimore, and 66 independent AIMs for Chicago). Among case-control samples, the mean African ancestry was significantly higher in asthmatics than in non-asthmatics (82.0±14.0% vs. 77.8±18.1%, mean difference 4.2% [95% confidence interval (CI):2.0–6.4], p<0.0001). This association remained significant after adjusting for potential confounders (odds ratio: 4.55, 95% CI: 1.69–12.29, p = 0.003). African ancestry failed to show an association with asthma exacerbations (p = 0.965) using a model based on longitudinal data of the number of exacerbations followed over 1.5 years.

Conclusions/Significance

These data replicate previous findings indicating that African ancestry constitutes a risk factor for asthma and suggest that elevated asthma rates in African Americans can be partially attributed to African genetic ancestry.     

Preference for an invasive fruit trumps fruit abundance in selection by an introduced bird in the Society Islands, French Polynesia

Introduced plants with fleshy fruit can alter the dietary decisions of frugivorous birds in their novel ranges by producing fruit of higher quality or by producing fruit in greater abundance. We used fruit choice experiments with wild-caught captive Red-vented Bulbuls (Pycnonotus cafer) on the tropical Pacific island of Moorea, French Polynesia, to determine whether this bird prefers the fruit of a highly invasive tree (Miconia calvescens) over three other fruit (one alien, two native) and to determine whether birds would eat less preferred fruit when it was more abundant than preferred fruit. Birds showed consistent preferences, and chose M. calvescens more than any other species. Birds selected more abundant fruit first when a single species was presented. However, when both fruit species and abundance were modified simultaneously, patterns of preference for particular species remained intact while the response to abundance disappeared. Results imply that dietary preferences are more important than small-scale variations in abundance for fruit selection. The strong preference for M. calvescens suggests that Bulbuls will select the fruit even in habitats where it is rare.     

Kinetic studies of catechol-o-methyltransferase from the brain of the African catfish,Clarias gariepinus

• 1.1. Optimum in vitro conditions, and kinetics of the enzyme catechol-O-methyltransferase from the brain of the male African catfish were studied.
• 2.2. A saturated level for S-adenosylmethionine, as methyldonor, and magnesium as cofactor was reached at 5 μM and 10 mM, respectively.
• 3.3. The addition of ascorbic acid, as an antioxidant, and tranylcypromine, as a MAO inhibitor, was not necessary, during incubations with fore-brain homogenates.
• 4.4. Kinetic analysis of the methylation of catecholestrone, catecholestradiol and dopamine showed K_m values of 1.2, 0.6 and 0.5 μM, respectively.
• 5.5. The affinity of the catecholsubstrates for the enzyme catechol-O-methyltransferase is much higher in the brain of the African catfish than in tissues of mammals.

     

Viral Glycoprotein Complex Formation,Essential Function and Immunogenicity in the Guinea Pig Model for Cytomegalovirus

Development of a cytomegalovirus (CMV) vaccine is a major public health priority due to the risk of congenital infection. A key component of a vaccine is thought to be an effective neutralizing antibody response against the viral glycoproteins necessary for cell entry. Species specificity of human CMV (HCMV) precludes direct studies in an animal model. The guinea pig is the only small animal model for congenital cytomegalovirus infection. Analysis of the guinea pig CMV (GPCMV) genome indicates that it potentially encodes homologs to the HCMV glycoproteins (including gB, gH, gL, gM, gN and gO) that form various cell entry complexes on the outside of the virus: gCI (gB); gCII (gH/gL/gO); gCIII (gM/gN). The gB homolog (GP55) has been investigated as a candidate subunit vaccine but little is known about the other homolog proteins. GPCMV glycoproteins were investigated by transient expression studies which indicated that homolog glycoproteins to gN and gM, or gH, gL and gO were able to co-localize in cells and generate respective homolog complexes which could be verified by immunoprecipitation assays. ELISA studies demonstrated that the individual complexes were highly immunogenic in guinea pigs. The gO (GP74) homolog protein has 13 conserved N-glycosylation sites found in HCMV gO. In transient expression studies, only the glycosylated protein is detected but in virus infected cells both N-glycosylated and non-glycosylated gO protein were detected. In protein interaction studies, a mutant gO that lacked N-glycosylation sites had no impact on the ability of the protein to interact with gH/gL which indicated a potential alternative function associated with these sites. Knockout GPCMV BAC mutagenesis of the respective glycoprotein genes (GP55 for gB, GP75 for gH, GP115 for gL, GP100 for gM, GP73 for gN and GP74 for gO) in separate reactions was lethal for virus regeneration on fibroblast cells which demonstrated the essential nature of the GPCMV glycoproteins. The gene knockout results were similar to HCMV, except in the case of the gO homolog, which was non-essential in epithelial tropic virus but essential in lab adapted GPCMV. Overall, the findings demonstrate the similarity between HCMV and GPCMV glycoproteins and strengthen the relevance of this model for development of CMV intervention strategies.     

Carbohydrate-Binding Non-Peptidic Pradimicins for the Treatment of Acute Sleeping Sickness in Murine Models

Current treatments available for African sleeping sickness or human African trypanosomiasis (HAT) are limited, with poor efficacy and unacceptable safety profiles. Here, we report a new approach to address treatment of this disease based on the use of compounds that bind to parasite surface glycans leading to rapid killing of trypanosomes. Pradimicin and its derivatives are non-peptidic carbohydrate-binding agents that adhere to the carbohydrate moiety of the parasite surface glycoproteins inducing parasite lysis in vitro. Notably, pradimicin S has good pharmaceutical properties and enables cure of an acute form of the disease in mice. By inducing resistance in vitro we have established that the composition of the sugars attached to the variant surface glycoproteins are critical to the mode of action of pradimicins and play an important role in infectivity. The compounds identified represent a novel approach to develop drugs to treat HAT.     

Increased geographic sampling reveals considerable new genetic diversity in the morphologically conservative African Pygmy Mice (Genus Mus; Subgenus Nannomys)

African endemic pygmy mice (Genus Mus; sub-genus Nannomys) have considerable economic and public health significance, and some species exhibit novel sex determination systems, making accurate knowledge of their phylogenetics and distribution limits important. This phylogenetic study was based on the mitochondrial control region and cytochrome b gene, for which a substantial body of published data was available. Study specimens were sourced from eight previously unsampled or poorly sampled countries, and include samples morphologically identified as Mus bufo, M. indutus, M. callewaerti, M. triton and M. neavei. These analyses increase the known genetic diversity of Nannomys from 65 to 102 haplotypes; at least 5 unassigned haplotypes are distinguished by potentially species-level cytochrome b genetic distances. The monophyly of Nannomys is supported. Mus musculoides, M. callewaerti, M. indutus, M. bufo, M. haussa, M. mattheyi, M. baoulei and M. sorella are supported as discrete species. The range of M. indutus is extended to include Botswana. M. setulosus and M. minutoides appear to be species complexes. A south and east African M. minutoides clade was defined and includes 8 new haplotypes out of 15. M. setulosus sensu lato includes M. setulosus sensu stricto and a strongly-supported M. bufo clade. Two samples, morphologically identified as M. triton and M. neavei, fall within the M. minutoides clade.     

Carbohydrate-Dependent Binding of Langerin to SodC,a Cell Wall Glycoprotein of Mycobacterium leprae

Langerhans cells participate in the immune response in leprosy by their ability to activate T cells that recognize the pathogen, Mycobacterium leprae, in a langerin-dependent manner. We hypothesized that langerin, the distinguishing C-type lectin of Langerhans cells, would recognize the highly mannosylated structures in pathogenic Mycobacterium spp. The coding region for the extracellular and neck domain of human langerin was cloned and expressed to produce a recombinant active trimeric form of human langerin (r-langerin). Binding assays performed in microtiter plates, by two-dimensional (2D) Western blotting, and by surface plasmon resonance demonstrated that r-langerin possessed carbohydrate-dependent affinity to glycoproteins in the cell wall of M. leprae. This lectin, however, yielded less binding to mannose-capped lipoarabinomannan (ManLAM) and even lower levels of binding to phosphatidylinositol mannosides. However, the superoxide dismutase C (SodC) protein of the M. leprae cell wall was identified as a langerin-reactive ligand. Tandem mass spectrometry verified the glycosylation of a recombinant form of M. leprae SodC (rSodC) produced in Mycobacterium smegmatis. Analysis of r-langerin affinity by surface plasmon resonance revealed a carbohydrate-dependent affinity of rSodC (equilibrium dissociation constant [K_D] = 0.862 μM) that was 20-fold greater than for M. leprae ManLAM (K_D = 18.69 μM). These data strongly suggest that a subset of the presumptively mannosylated M. leprae glycoproteins act as ligands for langerin and may facilitate the interaction of M. leprae with Langerhans cells.     

The African species Megantereon whitei from the Early Pleistocene of Monte Argentario (South Tuscany,Central Italy)

A partial skull and articulated postcranial elements of Megantereon whitei have been recorded during the 1950s from a karst deposit in the Monte Argentario area (Grosseto). These fossils recently became available for study. The bones are quite well preserved, included in a hard reddish matrix with calcareous clasts. The fossil is part of a faunal assemblage referred to the Late Villafranchian (Early Pleistocene). M. whitei from Monte Argentario is characterised by elongated upper canines, stronger than those of the Upper Valdarno (Tuscany) specimens and similar to the Pirro Nord (Apulia) fossil. The manus is robust, the first phalanges are quite long in comparison to the metacarpals. The Early Pleistocene European M. whitei represents an African element that took part in the faunal dispersal from Africa to Europe that occurred at the Plio-Pleistocene transition.