Carboxyamidation of Purine Nucleosides: New Secondary 8-Carboxamidopurine Nucleosides

Abstract

Palladium catalyzed carboxyamidation at the 8-position of 8-bromoadenosine and 8-bromoguanosine nucleosides is a versatile reaction, which allows primary, secondary, heterocyclic, aromatic mine and amino acids to be incorporated into purine nucleosides.     

Enhancement of phenolic compounds oxidation using laccase from Trametes versicolor in a microreactor

Laccases catalyse the oxidation of a wide range of substrates by a radical-catalyzed reaction mechanism, with a corresponding reduction of oxygen to water in a four-electron transfer process. Due to that, laccases are considered environmentally friendly enzymes, and lately there has been great interest in their use for the transformation and degradation of phenolic compounds. In this work, enzymatic oxidation of catechol and L-DOPA using commercial laccase from Trametes versicolor was performed, in continuously operated microreactors. The main focus of this investigation was to develop a new process for phenolic compounds oxidation, by application of microreactors. For a residence time of 72 s and an inlet oxygen concentration of 0.271 mmol/dm³, catechol conversion of 41.3% was achieved, while approximately the same conversion of L-DOPA (45.0%) was achieved for an inlet oxygen concentration of 0.544 mmol/dm³. The efficiency of microreactor usage for phenolic compounds oxidation was confirmed by calculating the oxidation rates; in the case of catechol oxidation, oxidation rates were in the range from 76.101 to 703.935 g/dm³/d (18–167 fold higher, compared to the case in a macroreactor). To better describe the proposed process, kinetic parameters of catechol oxidation were estimated, using data collected from experiments performed in a microreactor. The maximum reaction rate estimated in microreactor experiments was two times higher than one estimated using the initial reaction rate method from experiments performed in a cuvette. A mathematical model of the process was developed, and validated, using data from independent experiments.     

Field performance of Xa21 transgenic indica rice (Oryza sativa L.), IR72

Based on the characterization of the resistance phenotype and molecular analysis, several homozygous lines carrying Xa21 against the bacterial blight (BB) pathogen were obtained from previously transformed indica rice, IR72. The homozygous line, T103-10, with the best phenotype and seed-setting, was repeatedly tested under normal field conditions to evaluate its levels of resistance to the BB pathogen in Wuhan, China, in 1998 and 1999. The isolates of Xanthomonas oryzae pv oryzae (Xoo) used in this experiments were PXO61, PXO79, PXO99 and PXO112 isolated from the Philippines, T2 isolated from Japan, and Zhe173 isolated from China. The results demonstrated that the transgenic homozygous line expressed the same resistance spectrum, but with a shorter lesion length to each inoculated isolates as the lesion length of the Xa21 donor line IRBB21. The non-transformed control IR72 carrying Xa4 was resistant to PXO61, PXO112, Zhe173 and T2, but susceptible to PXO99 and PXO79. The negative control variety IR24 was susceptible to all isolates under field conditions. The results demonstrated clearly that the Xa21 transgene led to an excellent field performance of the introduced bacterial blight resistance trait on the recipient plants. The yield performance of this transgenic homozygous line, T103-10, is comparable with that of the control under field conditions. Received: 2 August 1999 / Accepted: 3 November 1999     

Phylogeny of some systematically uncertain urostyloids--Apokeronopsis, Metaurostylopsis, Thigmokeronopsis (Ciliophora, Stichotrichia) estimated with small subunit rRNA gene sequence information: discrepancies and agreements with morphological data

The small subunit rRNA (SSrRNA) genes of seven species of urostyloids representing four genera were sequenced. These were: Apokeronopsis crassa, A. bergeri, Anteholosticha sp-QD-1, Metaurostylopsis sp-QD-1, M. sp-QD-2, M. sp-QD-3 and Thigmokeronopsis sp-QD-1. Gene trees were constructed in order to investigate their phylogenetic relationships. The results indicate that: (1) Apokeronopsis, Thigmokeronopsis and Metaurostylopsis form a well-supported, clearly isolated, monophyletic group; (2) Metaurostylopsis species analysed consistently group together indicating that it is a well-outlined genus; (3) the validity of the genus Apokeronopsis is supported; (4) the separation of Holosticha and Anteholosticha is supported although Anteholosticha species exhibit a high molecular diversity; (5) Pseudokeronopsis and Thigmokeronopsis, which have been considered closely related, may not share a recent common ancestor, casting doubt on the monophyly of the family Pseudokeronopsidae.     

Individual variations in arsenic metabolism in Vietnamese: the association with arsenic exposure and GSTP1 genetic polymorphism

We investigated the association of As exposure and genetic polymorphism in glutathione S-transferase π1 (GSTP1) with As metabolism in 190 local residents from the As contaminated groundwater areas in the Red River Delta, Vietnam. Total As concentrations in groundwater ranged from <0.1 to 502 μg l(-1). Concentrations of dimethylarsinic acid (DMA(V)), monomethylarsonic acid (MMA(V)), and arsenite (As(III)) in human urine were positively correlated with total As levels in the groundwater, suggesting that people in these areas may be exposed to As through the groundwater. The concentration ratios of urinary As(III)/arsenate (As(V)) and MMA(V)/inorganic As (IA; As(III) + As(V))(M/I), which are indicators of As metabolism, increased with the urinary As level. Concentration and proportion of As(III) were high in the wild type of GSTP1 Ile105Val compared with the hetero type, and these trends were more pronounced in the higher As exposure group (>56 μg l(-1) creatinine in urine), but not in the lower exposure group. In the high As exposure group, As(III)/As(V) ratios in the urine of wild type of GSTP1 Ile105Val were significantly higher than those of the hetero type, while the opposite trend was observed for M/I. These results suggest that the excretion and metabolism of IA may depend on both the As exposure level and the GSTP1 Ile105Val genotype.     

Physiologic diversity and development of intrinsically photosensitive retinal ganglion cells

Intrinsically photosensitive retinal ganglion cells (ipRGCs) mediate numerous nonvisual phenomena, including entrainment of the circadian clock to light-dark cycles, pupillary light responsiveness, and light-regulated hormone release. We have applied multielectrode array recording to characterize murine ipRGCs. We find that all ipRGC photosensitivity is melanopsin dependent. At least three populations of ipRGCs are present in the postnatal day 8 (P8) murine retina: slow onset, sensitive, fast off (type I); slow onset, insensitive, slow off (type II); and rapid onset, sensitive, very slow off (type III). Recordings from adult rd/rd retinas reveal cells comparable to postnatal types II and III. Recordings from early postnatal retinas demonstrate intrinsic light responses from P0. Early light responses are transient and insensitive but by P6 show increased photosensitivity and persistence. These results demonstrate that ipRGCs are the first light-sensitive cells in the retina and suggest previously unappreciated diversity in this cell population.     

Two CheW coupling proteins are essential in a chemosensory pathway of Borrelia burgdorferi

In the model organism Escherichia coli, the coupling protein CheW, which bridges the chemoreceptors and histidine kinase CheA, is essential for chemotaxis. Unlike the situation in E. coli, Borrelia burgdorferi, the causative agent of Lyme disease, has three cheW homologues (cheW₁, cheW₂ and cheW₃). Here, a comprehensive approach is utilized to investigate the roles of the three cheWs in chemotaxis of B. burgdorferi. First, genetic studies indicated that both the cheW₁ and cheW₃ genes are essential for chemotaxis, as the mutants had altered swimming behaviours and were non‐chemotactic. Second, immunofluorescence and cryo‐electron tomography studies suggested that both CheW₁ and CheW₃ are involved in the assembly of chemoreceptor arrays at the cell poles. In contrast to cheW₁ and cheW₃, cheW₂ is dispensable for chemotaxis and assembly of the chemoreceptor arrays. Finally, immunoprecipitation studies demonstrated that the three CheWs interact with different CheAs: CheW₁ and CheW₃ interact with CheA₂ whereas CheW₂ binds to CheA₁. Collectively, our results indicate that CheW₁ and CheW₃ are incorporated into one chemosensory pathway that is essential for B. burgdorferi chemotaxis. Although many bacteria have more than one homologue of CheW, to our knowledge, this report provides the first experimental evidence that two CheW proteins coexist in one chemosensory pathway and that both are essential for chemotaxis.     

Molecular phylogeny of Nothoholosticha (Protozoa,Ciliophora, Urostylida) and systematic relationships of the Holosticha-complex

The marine ciliate Nothoholosticha is characterized by having a Holosticha-like ciliature pattern but without migratory frontoterminal cirri. However, its systematic position and its relationship to other members of the Holosticha-complex have not yet been resolved. In order to gain deeper insights into these relationships, the small subunit rRNA (SSrRNA) gene and the rRNA internal transcribed spacer and 5.8SrRNA (ITS1–5.8S-ITS2) region of two marine Anteholosticha species, as well as the ITS1–5.8S-ITS2 region of N. fasciola, were sequenced, and molecular trees (BI, ML, NJ and MP trees) were constructed. Although our analyses failed to conclusively resolve the phylogeny of this assemblage, certain conclusions could be drawn. Firstly, Nothoholosticha is a valid genus that is more closely related to Pseudokeronopsis than to other Holosticha-complex genera. Secondly, sequence analyses and phylogenetic trees of several Anteholosticha species revealed a high molecular diversity, which does not support the monophyly of this genus. Thirdly, the current assignment of certain well-known genera, e.g. Holosticha, Anteholosticha, Apokeronopsis, Parabirojimia, Psammomitra, Diaxonella, Metaurostylopsis and Thigmokeronopsis, to the families Bakuellidae (sensu Berger 2006), Urostylidae (sensu Berger 2006) or Holostichidae (sensu Berger 2006) is challenged by the molecular data presented here. And fourthly, the families Holostichidae and Pseudokeronopsidae (sensu Lynn 2008) are probably paraphyletic, and their systematic assignments await further evaluation.