In vitro susceptibility of human Blastocystis subtypes to simeprevir

Introduction and aimBlastocystis is a common enteric parasite, having a worldwide distribution. Many antimicrobial agents are effective against it, yet side effects and drug resistance have been reported. Thus, ongoing trials are being conducted for exploring anti-Blastocystis alternatives. Proteases are attractive anti-protozoal drug targets, having documented roles in Blastocystis. Serine proteases are present in both hepatitis C virus and Blastocystis. Since drug repositioning is quite trendy, the in vitro efficacy of simeprevir (SMV), an anti-hepatitis serine protease inhibitor, against Blastocystis was investigated in the current study.MethodsStool samples were collected from patients, Alexandria, Egypt. Concentrated stools were screened using direct smears, trichrome, and modified Ziehl-Neelsen stains to exclude parasitic co-infections. Positive stool isolates were cultivated, molecularly subtyped for assessing the efficacy of three SMV doses (100,150, and 200 μg/ml) along 72 hours (h), on the most common subtype, through monitoring parasite growth, viability, re-culture, and also via ultrastructure verification. The most efficient dose and duration were later tested on other subtypes.ResultsResults revealed that Blastocystis was detected in 54.17% of examined samples. Molecularly, ST3 predominated (62%), followed by ST1 (8.6%) and ST2 (3.4%). Ascending concentrations of SMV progressively inhibited growth, viability, and re-culture of treated Blastocystis, with a non-statistically significant difference when compared to the therapeutic control metronidazole (MTZ). The most efficient dose and duration against ST3 was 150 µg/ml for 72 h. This dose inhibited the growth of ST3, ST1, and ST2 with percentages of 95.19%, 94.83%, and 94.74%, successively and viability with percentages of 98.30%, 98.09%, and 97.96%, successively. This dose abolished Blastocystis upon re-culturing. Ultra-structurally, SMV induced rupture of Blastocystis cell membrane leading to necrotic death, versus the reported apoptotic death caused by MTZ. In conclusion, 150 µg/ml SMV for 72 h proved its efficacy against ST1, ST2, and ST3 Blastocystis, thus sparing the need for pre-treatment molecular subtyping in developing countries.     

The profile of free amino acids in latent fingerprint of healthy and beta-thalassemic volunteers

The aim of the present work is to apply a non-invasive test, using thumb fingerprint residue analysis, for detection of beta-thalassemia (β-Thal). The relative percentages of free amino acids (AA) in the latent fingerprint of β-Thal patients and healthy subjects were compared. The sample included 24 β-Thal patient and 24 healthy subjects, aged 5–10 years old. Twenty-three AA plus ammonia were analyzed by a sensitive high-performance liquid chromatographic method with fluorescence detection. The profile of AA was calculated based on the percentage of relative amount of each AA to serine (Ser) as it found to be the predominant peak. The statistical and chromatographic profiles of β-Thal patients were characterized by significant decrease of ornithine, lysine, and zero tyrosine, with significant increase of ammonia, and proline. Other amino acids that exist in low ratios were estimated statistically for significance changes. The relative percentages of each AA of healthy subjects were approximately constant. For this reason, these mentioned AA were assigned as major fingerprint markers of β-Thal.     

Phylogenetic diversity and ecological features in the Egyptian flora

Until fairly recently, regional-scale ecological and evolutionarypatterns have tended to be ignored as conservation efforts have been concernedwith species and their habitats. Here we compare frequencies in the Egyptianflora of particular rank sizes (order, family and genus) with patterns ofspecies abundance (classified as very rare, rare, common, or very common) and anarray of life-history attributes. The angiosperm flora of Egypt is representedby 2446 taxa (2088 species), including taxa in 10 subclasses, 51 orders, 120families, and 742 genera. A high degree of monotypism was observed: four ordersare monotypic (each existing as single species), and have very rare overallabundances; 30 families are monotypic (17 of which are very rare or rare); and 354genera are monotypic (over 70% of which are very rare or rare). Fourteenfamilies (in particular the Resedaceae and Zygophyllaceae) have at leastone-fifth of their global species represented in the Egyptian flora. Introducedspecies in general, and tree, aquatic herb and liana life forms all are especially well represented among monotypic genera. Native taxa are highlyrepresented among rare and very rare abundance classes, while introduced taxadid not differ significantly in their abundance patterns, compared to overallflora values. Few large genera (>20 spp.) occur in the flora, with mostspecies concentrated in genera containing 8–19 species per genus.Similarly, few families were highly speciose. Annual and herbaceous species weresignificantly over-represented, mainly among large, speciose genera andfamilies. However, perennials, trees, shrubs, aquatic herbs, lianas and parasiticspecies were found mainly in families and genera having very few taxa.Life-history attributes may have important implications to speciation rates.Taxonomically based results, involving abundances and life-history attributes,are discussed in the context of biodiversity and conservation.