Centrosomal localization of cyclins E and A: Structural similarities and functional differences

Recent identification of the modular CLS motifs responsible for cyclins A and E localization on centrosomes has revealed a tight linkage between the nuclear and centrosomal cycles. These G₁/S cyclins must localize on the centrosome in order for DNA replication to occur in the nucleus, whereas essential DNA replication factors also function on the centrosome to prevent centrosome overduplication. Both events are dependent on the presence of an intact CLS within each cyclin. Here we compare the cyclins A and E CLSs at the structural and functional levels and identify a new cyclin A CLS mutant that disrupts all CLS functions and reduces the affinity of cyclin A for Cdk2. Analysis of interactions of the CLS motif within the cyclin molecules highlights the importance of the cyclin CBOX1 region for Cdk2 binding.Key words: cyclin A, cyclin E, Cdk2, centrosome, CLS, PSTAIRE, DNA synthesis     

A Canadian Critical Care Trials Group project in collaboration with the international forum for acute care trialists - Collaborative H1N1 Adjuvant Treatment pilot trial (CHAT): study protocol and design of a randomized controlled trial

Background

The Vaccine Assessment using Linked Data (VALiD) trial compared opt-in and opt-out parental consent for a population-based childhood vaccine safety surveillance program using data linkage. A subsequent telephone interview of all households enrolled in the trial elicited parental intent regarding the return or non-return of reply forms for opt-in and opt-out consent. This paper describes the rationale for the trial and provides an overview of the design and methods.

Methods/Design

Single-centre, single-blind, randomised controlled trial (RCT) stratified by firstborn status. Mothers who gave birth at one tertiary South Australian hospital were randomised at six weeks post-partum to receive an opt-in or opt-out reply form, along with information explaining data linkage. The primary outcome at 10 weeks post-partum was parental participation in each arm, as indicated by the respective return or non-return of a reply form (or via telephone or email response). A subsequent telephone interview at 10 weeks post-partum elicited parental intent regarding the return or non-return of the reply form, and attitudes and knowledge about data linkage, vaccine safety, consent preferences and vaccination practices. Enrolment began in July 2009 and 1,129 households were recruited in a three-month period. Analysis has not yet been undertaken. The participation rate and selection bias for each method of consent will be compared when the data are analysed.

Discussion

The VALiD RCT represents the first trial of opt-in versus opt-out consent for a data linkage study that assesses consent preferences and intent compared with actual opting in or opting out behaviour, and socioeconomic factors. The limitations to generalisability are discussed.

Trial registration

Australian New Zealand Clinical Trials Registry ACTRN12610000332022     

Using caching and optimization techniques to improve performance of the Ensembl website

Background  

The Ensembl web site has provided access to genomic information for almost 10 years. During this time the amount of data available through Ensembl has grown dramatically. At the same time, the World Wide Web itself has become a dramatically more important component of the scientific workflow and the way that scientists share and access data and scientific information.     

Effects of repeated acupuncture action on pain sensitivity and beta-endorphin contents of the hypothalamus and midbrain of the rat

Changes in latencies (L) of nociceptive (NC) reactions of hot plate (HP) tail flick (TF) tests and dynamics of beta-endorphin (beta-ED) in hypothalamus and midbrain were studied in albino rats receiving 15 sessions of acupuncture (A) (20 min daily in bilateral Ho-Ku). It was shown that repeated A facilitated prolongation of LHP and LTF as compared to baseline and control levels. A decrease of the anti-NC systems capacity for activation in response to each A stimulation was observed. The dynamics of beta-endorphin in hypothalamus showed an opposite tendency as compared to that in midbrain. It is suggested that such a correlation of beta-ED activities in midbrain and hypothalamus might be one of the reasons for anti-NC mechanisms adaptation to A.     

Thermovision monitoring of neurografts in the rat

By thermovision technique and images digital processing, thermal fields and reactions of dorsal cortical surface of 15 white rats were studied through the intact skull in various times after transplantation of the embryonal visual cortex, amygdala and hippocampus into the visual cortex. After operation significant thermoasymmetries arise outside transplantation area which in early periods (2-15 days) are dynamic, and in 5-5.5 months become stabilized. At successful implantation, transplantat area in background state in late terms is not thermodistinct, and in pseudo-transplanted animals--is cooled. In transplantat area, a changed thermoreaction to light stimulation is recorded, and in early terms it is connected with thermal flow from neighbouring cortical sites.     

Demographic consequences of nestbox use for Red‐footed Falcons Falco vespertinus in Central Asia

Nestbox programmes are frequently implemented for the conservation of cavity‐nesting birds, but their effectiveness is rarely evaluated in comparison with birds not using nestboxes. In the European Palaearctic, Red‐footed Falcon Falco vespertinus populations are both of high conservation concern and are strongly associated with nestbox programmes in heavily managed landscapes. We used a 21‐year monitoring dataset collected on 753 nesting attempts by Red‐footed Falcons in unmanaged natural or semi‐natural habitats to provide basic information on this poorly known species; to evaluate long‐term demographic trends within this population; and to evaluate response of demographic parameters of Red‐footed Falcons to environmental factors including use of nestboxes. We observed significant differences among years in laying date, offspring loss and numbers of fledglings produced, but not in egg production. Of these four parameters, offspring loss and, to a lesser extent, number of fledglings exhibited directional trends over time. Variation in laying date and in numbers of eggs were not well explained by any one model of environmental factors, but instead by combinations of models, each with informative terms for nest type. Nevertheless, laying in nestboxes occurred 2.10 ± 0.70 days earlier than in natural nests. In contrast, variation in both offspring loss and numbers of fledglings produced were fairly well explained by a single model including terms for nest type, nest location and an interaction between the two parameters (65 and 81% model weights, respectively), with highest offspring loss in nestboxes on forest edges. Because, for other species, earlier laying dates are associated with more fit individuals, this interaction highlighted a possible ecological trap, whereby birds using nestboxes on forest edges lay eggs earlier but suffer greater offspring loss and produce lower numbers of fledglings than do those in other nesting settings. If nestboxes increase offspring loss for Red‐footed Falcons in heavily managed landscapes where populations are at greater risk, or for the many other species of rare or endangered birds supported by nestbox programmes, these processes could have important demographic and conservation consequences.     

NMR relaxation parameters of methyl groups as a tool to map the interfaces of helix–helix interactions in membrane proteins

A number of recent advances in the field of magic-angle-spinning (MAS) solid-state NMR have enabled its application to a range of biological systems of ever increasing complexity. To retain biological relevance, these samples are increasingly studied in a hydrated state. At the same time, experimental feasibility requires the sample preparation process to attain a high sample concentration within the final MAS rotor. We discuss these considerations, and how they have led to a number of different approaches to MAS NMR sample preparation. We describe our experience of how custom-made (or commercially available) ultracentrifugal devices can facilitate a simple, fast and reliable sample preparation process. A number of groups have since adopted such tools, in some cases to prepare samples for sedimentation-style MAS NMR experiments. Here we argue for a more widespread adoption of their use for routine MAS NMR sample preparation.     

A non-invasive genetic evaluation of population size,natal philopatry,and roosting behavior of non-breeding eastern imperial eagles (<Emphasis Type="Italic">Aquila heliaca</Emphasis>) in central Asia

Roughly one-third of all eagle species are considered to be threatened or endangered, but the ecology of most eagles remains poorly understood. While the pronounced territoriality of breeding adults facilitates behavioral studies, the demography of non-breeding individuals (pre-adults and non-territorial floaters) is almost completely unknown. Traditionally, limited data on pre-adult and floater movement come from wing-tagging and/or telemetry studies. As an alternative to these methods we used genetic analyses of non-invasively collected feathers to investigate the population biology of non-breeding eastern imperial eagles (Aquila heliaca) in Kazakhstan. Microsatellite profiles of shed feathers indicate that eastern imperial eagles roost communally with other raptors. Furthermore, roosts are large and dynamic: 287 non-breeding eastern imperial eagles were detected in our sample, and a mark-recapture analysis estimated the total number to be 308 ± 8. The natal origins of these individuals were investigated by comparing their microsatellite profiles to those available for >90% of the eastern imperial eagle chicks hatched at the study site over the six previous breeding seasons. Only 4% of the individuals genetically matched a chick, suggesting that the reserve may serve as a critical refugium for pre-adults and itinerant floaters. Feathers have long been recognized as a suitable source of DNA, but few studies have used wide-scale, non-invasive collections of feathers (>1,000 samples) to investigate fundamental aspects of avian biology. Our research demonstrates that non-invasive genetic analyses of feathers can be used to evaluate population size, natal philopatry, and local movements of birds that are difficult to study using traditional means.     

A 76-bp Deletion in the Mip Gene Causes Autosomal Dominant Cataract in Hfi Mice

Hfi is a dominant cataract mutation where heterozygotes show hydropic lens fibers and homozygotes show total lens opacity. The Hfi locus was mapped to the distal part of mouse chromosome 10 close to the major intrinsic protein (Mip), which is expressed only in cell membranes of lens fibers. Molecular analysis of Mip revealed a 76-bp deletion that resulted in exon 2 skipping in Mip mRNA. In Hfi/Hfi this deletion resulted in a complete absence of the wildtype Mip. In contrast, Hfi/+ animals had the same amount of wildtype Mip as +/+. Results from pulse–chase expression studies excluded hetero-oligomerization of wildtype and mutant Mip as a possible mechanism for cataract formation in the Hfi/+. We propose that the cataract phenotype in the Hfi heterozygote mutant is due to a detrimental gain of function by the mutant Mip resulting in either cytotoxicity or disruption in processing of other proteins important for the lens. Cataract formation in the Hfi/Hfi mouse is probably a combined result of both the complete loss of wildtype Mip and a gain of function of the mutant Mip.     

Experimental research on the cellular interactions during hemopoiesis

We present a review of experimental studies performed at the Laboratory of Histogenesis of the Institute of Developmental Biology, Russian Academy of Sciences, on the problem of cell interactions during hemopoiesis. Special attention has been given to original experimental models, such as production of hemopoietic foci on underlayers of fibroblasts encapsulating a foreign body in the peritoneal cavity of rodents (after intraperitoneal transplantation of hemopoietic cells) and repopulation of ectopic hemopoietic territories under the kidney capsule of mice by syngeneic or xenogeneic hemopoietic cells. We describe the competitive interactions of genetically different hemopoietic cells after the transplantation of their mixtures to irradiated mice (multicomponent radiation chimeras). Xenogeneic and multicomponent chimeras have also been obtained in long-term bone marrow culture. We have examined characteristics of hemopoiesis on stromal cell underlayers produced by cells of various origins in vitro and then transplanted into the peritoneal cavity of irradiated mice. We discuss the results obtained and possible mechanisms of these phenomena.