ABSTRACT
Processes catalyzed by enzymes offer numerous advantages over chemical methods although in many occasions the stability of the biocatalysts becomes a serious concern. Traditionally, synthesis of nucleosides using poorly water-soluble purine bases, such as guanine, xanthine, or hypoxanthine, requires alkaline pH and/or high temperatures in order to solubilize the substrate. In this work, we demonstrate that the 2'-deoxyribosyltransferase from Leishmania mexicana (LmPDT) exhibits an unusually high activity and stability under alkaline conditions (pH 8-10) across a broad range of temperatures (30-70 °C) and ionic strengths (0-500 mM NaCl). Conversely, analysis of the crystal structure of LmPDT together with comparisons with hexameric, bacterial homologues revealed the importance of the relationships between the oligomeric state and the active site architecture within this family of enzymes. Moreover, molecular dynamics and docking approaches provided structural insights into the substrate-binding mode. Biochemical characterization of LmPDT identifies the enzyme as a type I NDT (PDT), exhibiting excellent activity, with specific activity values 100- and 4000-fold higher than the ones reported for other PDTs. Interestingly, LmPDT remained stable during 36 h at different pH values at 40 °C. In order to explore the potential of LmPDT as an industrial biocatalyst, enzymatic production of several natural and non-natural therapeutic nucleosides, such as vidarabine (ara A), didanosine (ddI), ddG, or 2'-fluoro-2'-deoxyguanosine, was carried out using poorly water-soluble purines. Noteworthy, this is the first time that the enzymatic synthesis of 2'-fluoro-2'-deoxyguanosine, ara G, and ara H by a 2'-deoxyribosyltransferase is reported.
Subject(s)
Leishmania mexicana/enzymology , Nucleosides/biosynthesis , Pentosyltransferases/metabolism , Purines/chemistry , Amino Acid Sequence , Biocatalysis , Cloning, Molecular , Computational Biology , Enzymes, Immobilized , Hydrogen-Ion Concentration , Leishmania mexicana/genetics , Pentosyltransferases/genetics , Protein Conformation , Sequence Alignment , Substrate Specificity , TemperatureABSTRACT
The mite Ornithonyssus bursa (Berlese) (Mesostigmata: Macronyssidae) is considered a poultry pest causing important infestations in chickens and it is considered a potential vector of arbovirus. Despite being considered a common parasite in wild birds, there is scarce published information about its potential hosts and effects on them. Here we present new bird hosts for O. bursa, assess the presence of Alphavirus, Flavivirus and Bunyavirus in mites from three host species, and discuss its potential impact on wild bird populations. We found O. bursa infecting five raptor and six passerine wild bird species. For nine of these species, this is the first record of infection by O. bursa. Although all analysed mites were negative for the examined arboviruses, the small sample size of mites does not allow further conclusions at the present moment. Because of the general nature of this ectoparasite, its presence in migratory long dispersal and endangered bird species, and the seropositivity for arboviruses in some of the species studied here, we consider it critical to assess the role of O. bursa and other ectoparasites as vectors and reservoirs of pathogens and as potential deleterious agents in wild bird populations.
Subject(s)
Bird Diseases/parasitology , Birds , Mite Infestations/veterinary , Mites/physiology , Mites/virology , Alphavirus/isolation & purification , Animals , Argentina/epidemiology , Bird Diseases/epidemiology , Female , Flavivirus/isolation & purification , Host-Parasite Interactions , Male , Mite Infestations/epidemiology , Mite Infestations/parasitology , Orthobunyavirus/isolation & purification , PrevalenceABSTRACT
This article documents the addition of 171 microsatellite marker loci and 27 pairs of single nucleotide polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Bombus pauloensis, Cephalorhynchus heavisidii, Cercospora sojina, Harpyhaliaetus coronatus, Hordeum vulgare, Lachnolaimus maximus, Oceanodroma monteiroi, Puccinia striiformis f. sp. tritici, Rhea americana, Salmo salar, Salmo trutta, Schistocephalus solidus, Sousa plumbea and Tursiops aduncus. These loci were cross-tested on the following species: Aquila heliaca, Bulweria bulwerii, Buteo buteo, Buteo swainsoni, Falco rusticolus, Haliaeetus albicilla, Halobaena caerulea, Hieraaetus fasciatus, Oceanodroma castro, Puccinia graminis f. sp. Tritici, Puccinia triticina, Rhea pennata and Schistocephalus pungitii. This article also documents the addition of 27 sequencing primer pairs for Puffinus baroli and Bulweria bulwerii and cross-testing of these loci in Oceanodroma castro, Pelagodroma marina, Pelecanoides georgicus, Pelecanoides urinatrix, Thalassarche chrysostoma and Thalassarche melanophrys.
Subject(s)
Databases, Genetic , Microsatellite Repeats , Polymorphism, Single Nucleotide , Ecology , Genetic MarkersABSTRACT
Se presentan 13 enfermos que recibieron neoadyuvancia como parte del tratamiento del CPNPC, excluído el tumor del vértice de pulmón. Siete correspondieron al E IIIa (5 por N2 con invasión extracapsular y 2 por T3) Y 6 AL IIIb (3 por N3 y 3 por T4). Recibieron radioterapia 2 enfermos, quimioterapia 6, y 5 la combinación de quimio y radioterapia. Dos casos presentaron progresión de enfermedad, 2 están aún en protocolo y 9 tuvieron respuesta favorable siendo operados, 7 se resecaron y a 2 se los consideró irresecables. Falleció un enfermo resecado al 6§ día con distres respiratorio. Se analizan las posibles causas de respuesta favorable. El índice de respuesta clínica fue 81.8 por ciento (9/11 casos), el índice de resección 77,77 por ciento (7/9) y el grado de esterilización histológica en los resecados 14,28 por ciento (1/7). Se concluye que un grupo significativo de pacientes con enfermedad locorregionalmente avanzada se pueden beneficiar con esta modalidad terapéutica, mejorando su calidad de vida. El aumento de la sobrevida aún no ha sido comprobado.
Subject(s)
Humans , Male , Middle Aged , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/mortality , Carcinoma, Non-Small-Cell Lung/radiotherapy , Carcinoma, Non-Small-Cell Lung/surgery , Chemotherapy, Adjuvant , Lung NeoplasmsABSTRACT
Se presentan 13 enfermos que recibieron neoadyuvancia como parte del tratamiento del CPNPC, excluído el tumor del vértice de pulmón. Siete correspondieron al E IIIa (5 por N2 con invasión extracapsular y 2 por T3) Y 6 AL IIIb (3 por N3 y 3 por T4). Recibieron radioterapia 2 enfermos, quimioterapia 6, y 5 la combinación de quimio y radioterapia. Dos casos presentaron progresión de enfermedad, 2 están aún en protocolo y 9 tuvieron respuesta favorable siendo operados, 7 se resecaron y a 2 se los consideró irresecables. Falleció un enfermo resecado al 6º día con distres respiratorio. Se analizan las posibles causas de respuesta favorable. El índice de respuesta clínica fue 81.8 por ciento (9/11 casos), el índice de resección 77,77 por ciento (7/9) y el grado de esterilización histológica en los resecados
