Methods for L-ribooligonucleotide sequence determination using LCMS

The ability to verify the sequence of a nucleic acid-based therapeutic is an essential step in the drug development process. The challenge associated with sequence identification increases with the length and nuclease resistance of the nucleic acid molecule, the latter being an important attribute of therapeutic oligonucleotides. We describe methods for the sequence determination of Spiegelmers, which are enantiomers of naturally occurring RNA with high resistance to enzymatic degradation. Spiegelmer sequencing is effected by affixing a label or hapten to the 5′-end of the oligonucleotide and chemically degrading the molecule in a controlled fashion to generate fragments that are then resolved and identified using liquid chromatography-mass spectrometry. The Spiegelmer sequence is then derived from these fragments. Examples are shown for two different Spiegelmers (NOX-E36 and NOX-A12), and the specificity of the method is shown using a NOX-E36 mismatch control

Psychiatric services utilization in completed suicides of a youth centres population

BACKGROUND: From a retrospective study of youth centres (YCs) and coroner's files, we investigated youths' history of medical service utilization who died by suicide. This is the second of two papers on YCs population, the first paper having shown that the rate of psychopathology was higher in the YCs population compared to the general adolescent population. METHODS: From 1995 to 2000, 422 youths, aged 18 years and younger, died as a result of suicide in Quebec. More than one-third received services from YCs at some point. Using the provincial physician payment and hospitalization database, we examined physical and psychiatric service utilization according to time intervals, as well as hospitalization for psychiatric reasons in the individuals' lifetime and in the year preceding suicide. Suicides were matched to living YCs youths for age, sex, and geographic area. YCs controls were then subdivided into two groups based on file information pertaining to the presence or absence of suicidal behavior or ideation. RESULTS: Compared to living YCs youths, suicides had a higher rate of psychiatric service utilization in the week, month, 90 days, and year preceding suicide, as well as higher levels of lifetime hospitalization for psychiatric reasons than controls with or without a history of suicidal behavior or ideation. We found that 28.3% YCs suicides made use of psychiatric services in the year preceding suicide. CONCLUSION: The rate of psychiatric service utilization by YCs youth suicides is substantially inferior to the needs of this population. Our study underscores the need for appropriate recognition of psychiatric and suicidal problems among YCs population by social and psycho-educational professionals. At the same time, it highlights the issues of general practitioners' risk identification, psychiatric referral and treatment. Our findings suggest the need for improved organization and coordination of psychiatric services to ameliorate treatment delivery

Variation in survival rates for the alpine marmot (Marmota marmota): effects of sex, age, year, and climatic factors

International audienceWe examined variation in annual survival rates in a population of alpine marmots (Marmota marmota) according to intrinsic (sex and age) and extrinsic (year and climate) factors. We tested predictions concerning (i) a sex effect in a monogamous non-dimorphic species, (ii) age structure of survival rates in a mesomammal, and (iii) the annual variability effect and the contribution of stochastic climatic factors, especially snow cover, frost, and rainfall. We used a 8-year dataset of 367 marmots that were livetrapped and marked in La Sassiere Nature Reserve in the French Alps between 1990 and 1997. Survival and recapture rates were modelled using recent developments in capture-recapture models. Sex had no effect on survival rates, which agrees with the predictions of sexual selection. Survival rates for young of the year (YOY, from weaning to first birthday) were, on average, lower than in the older age class. In the older age class, annual variation occurred that was strongly related to the intensity of autumn frost. By determining the soil temperature at the beginning of hibernation, this factor, though short-lived, could have determined the energetic cost of hibernation. Neither annual variation nor an environmental effect was detected in YOY despite a large sample size. Social thermoregulation could contribute to the stability of YOY survival rates. As infanticide was common after the immigration of a new dominant male, survival of YOY seemed to depend more on social events than on stochastic climatic ones

Extra-pair paternity in the monogamous Alpine marmot revealed by nuclear DNA microsatellite analysis

Times Cited: 58International audienceThe genetic parentage and pedigrees of 35 litters from 12 family groups of monogamous Alpine marmot (Marmota marmota) in the French Alps were analyzed using six hypervariable sequence repeat nuclear loci (microsatellites). All of the members of these family groups were sampled during a 5-year period. Hairs taken directly from animals served as a source of DNA for amplification of the loci. Our results indicate that the genetic mating system of the Alpine marmot is quite different from a strictly monogamous breeding system. Extra-pair paternity occurred in 11 of the 35 litters (31.4%). Of the 134 juveniles typed, 26 (19.4%) could be attributed to extra-pair copulation (EPC). We examine hypotheses which could explain the evolution of EPC and discuss the different patterns of extra-pair mating