31P NMR Studies of the Binding of the Oligonucleotide (Ap)3A to an Oligodeoxythymidylate Covalently Linked to an Acridine Derivative

Abstract

³¹P NMR was used to study the specific interaction of an oligodeoxynucleotide containing four thymines and covalently attached to an acridine derivative through its 3-phosphate [(Tp)₄(CH₂)₅Acr] with a complementary oligoribonucleotide (Ap)₃A.³¹P-¹H and ¹H-¹H chemical shift correlation spectroscopies were jointly used to provide the assignment of the phosphorus resonances. A downfield shift of two phosphorus resonances of (Tp)₄(CH₂)₅Acr and of two phosphorus resonances of (Ap)₄A was observed upon complex formation. The assignment of the phosphorus resonances which are downfield shifted allowed us to propose a model involving an equilibrium between several 1:1 complexes where the acridine ring is intercalated between different A.T base pairs.     

Whole-exome sequencing identifies mutations in GPR179 leading to autosomal-recessive complete congenital stationary night blindness

Congenital stationary night blindness (CSNB) is a heterogeneous retinal disorder characterized by visual impairment under low light conditions. This disorder is due to a signal transmission defect from rod photoreceptors to adjacent bipolar cells in the retina. Two forms can be distinguished clinically, complete CSNB (cCSNB) or incomplete CSNB; the two forms are distinguished on the basis of the affected signaling pathway. Mutations in NYX, GRM6, and TRPM1, expressed in the outer plexiform layer (OPL) lead to disruption of the ON-bipolar cell response and have been seen in patients with cCSNB. Whole-exome sequencing in cCSNB patients lacking mutations in the known genes led to the identification of a homozygous missense mutation (c.1807C>T [p.His603Tyr]) in one consanguineous autosomal-recessive cCSNB family and a homozygous frameshift mutation in GPR179 (c.278delC [p.Pro93Glnfs^∗57]) in a simplex male cCSNB patient. Additional screening with Sanger sequencing of 40 patients identified three other cCSNB patients harboring additional allelic mutations in GPR179. Although, immunhistological studies revealed Gpr179 in the OPL in wild-type mouse retina, Gpr179 did not colocalize with specific ON-bipolar markers. Interestingly, Gpr179 was highly concentrated in horizontal cells and Müller cell endfeet. The involvement of these cells in cCSNB and the specific function of GPR179 remain to be elucidated.     

RNA interference against animal viruses: how morbilliviruses generate extended diversity to escape small interfering RNA control

Viruses are serious threats to human and animal health. Vaccines can prevent viral diseases, but few antiviral treatments are available to control evolving infections. Among new antiviral therapies, RNA interference (RNAi) has been the focus of intensive research. However, along with the development of efficient RNAi-based therapeutics comes the risk of emergence of resistant viruses. In this study, we challenged the in vitro propensity of a morbillivirus (peste des petits ruminants virus), a stable RNA virus, to escape the inhibition conferred by single or multiple small interfering RNAs (siRNAs) against conserved regions of the N gene. Except with the combination of three different siRNAs, the virus systematically escaped RNAi after 3 to 20 consecutive passages. The genetic modifications involved consisted of single or multiple point nucleotide mutations and a deletion of a stretch of six nucleotides, illustrating that this virus has an unusual genomic malleability.     

Learning influences host choice in tsetse

A learning capacity for feeding is described in many insect species including vectors of diseases, but has never been reported in tsetse flies (Diptera, Glossinidae), the cyclic vectors of human (sleeping sickness) and animal trypanosomoses in Africa. Repeated feeding on the same host species by a disease vector is likely to increase the within-species disease-transmission risk, but to decrease it between species. An experiment with cattle and reptiles in a stable provides evidence that the species of host selected for the second blood meal in tsetse flies depends on the host encountered for the first blood meal when the between-meal interval is 2 days. This preference disappears when the between-meal interval is extended to 3 days. The energetic advantages of this acquired preference and its importance in trypanosomoses epidemiology are discussed.     

TRPM1 Is Mutated in Patients with Autosomal-Recessive Complete Congenital Stationary Night Blindness

Night vision requires signaling from rod photoreceptors to adjacent bipolar cells in the retina. Mutations in the genes NYX and GRM6, expressed in ON bipolar cells, lead to a disruption of the ON bipolar cell response. This dysfunction is present in patients with complete X-linked and autosomal-recessive congenital stationary night blindness (CSNB) and can be assessed by standard full-field electroretinography (ERG), showing severely reduced rod b-wave amplitude and slightly altered cone responses. Although many cases of complete CSNB (cCSNB) are caused by mutations in NYX and GRM6, in ∼60% of the patients the gene defect remains unknown. Animal models of human diseases are a good source for candidate genes, and we noted that a cCSNB phenotype present in homozygous Appaloosa horses is associated with downregulation of TRPM1. TRPM1, belonging to the family of transient receptor potential channels, is expressed in ON bipolar cells and therefore qualifies as an excellent candidate. Indeed, mutation analysis of 38 patients with CSNB identified ten unrelated cCSNB patients with 14 different mutations in this gene. The mutation spectrum comprises missense, splice-site, deletion, and nonsense mutations. We propose that the cCSNB phenotype in these patients is due to the absence of functional TRPM1 in retinal ON bipolar cells.     

Evaluation of crude annual parturition rate estimates in a small-holder African ruminant farming system

Many parameters have been proposed for evaluating livestock reproduction performances in tropical farming systems. In tropical free-ranged and small-holder systems, where reproduction cycles cannot be individually observed without expensive field surveys, one of these parameters is the average number of parturitions (h) expected by reproductive female, if the female spends the whole year in the herd. A frequent approach for estimating h is to use the ratio hc = m/T, so-called 'crude annual parturition rate', where m is the observed number of parturitions and T is the total time of presence of the reproductive females in the herd during the year. The bias encountered when h is estimated by hc was evaluated in this paper. Six methods of estimation were used, where T was the exact observed time of presence (hc1) or approximated by monthly, quarterly, half-yearly and yearly averages or final size of the reproductive herd size (hc2 to hc6). Data came from long-term follow-up of cattle and small-ruminant herds (with data recorded at animal level) in extensive agro-pastoral systems in Senegal. In general, h was correctly estimated by hc1. Nevertheless hc1 was sensitive to competing risks (e.g. deaths, sales and slaughtering of reproductive females) and was seriously biased when intensive withdrawals of females occurred before or during the parturition peak. Reliability of crude rates progressively decreased from hc2 to hc6, corresponding to the degradation of information used for approximating T. This decrease was much lower for cattle (for which all methods had acceptable reliability) than for small ruminants. Among the compared methods, the lower reliability was observed for hc6 that we do not recommend for small ruminants. Methods hc5 and hc6 are currently used in rapid cross-sectional retrospective surveys based on the recall of the farmers on the demographic events which occurred in the herd over the last past 12 months. The study has showed that such surveys and estimates hc5 and hc6 can generate seriously biased results. More globally, annual parturition rates can be highly variable depending on the 12-month periods considered. Annual parturition rates estimated on short-term data, even with precise herd follow-up surveys, must be considered cautiously.     

Modelling microbial processes (phyto- and bacterioplankton) in the Schelde estuary

Since about 10 years, studies have been conducted at the University of Brussels in modelling the microbiological processes affecting biogenic substances in the Schelde estuary and its watershed.The first model, a one dimensional redox model, simulated the longitudinal distribution of oxygen, nitrate, iron and manganese in relation to the observed bacterial heterotrophic activity. A model allowing calculation of bacterial activity from data on organic discharge was coupled to this model. It was completed by a model of phytoplanktonic development within the estuary. Finally, an idealized model of the hydrographical network, based on Horton analysis, is being established in order to calculate the quality of the water at the downward boundary of the estuary.Together, all these sub-models form a general model of the ecological working of the Schelde estuary, able to predict at least the general trends of the redox state, the organic carbon, mineral nitrogen, and chlorophyll-a concentrations as a function of distance to the sea, from the knowledge of geomorphological and meteorological data, along with informations concerning the distribution of anthropogenic discharges. This model therefore provides a powerfull tool for the rational management of the Schelde estuary.Research Associate of the Belgian National Scientific Research Funds.