All JNKs can kill, but nuclear localization is critical for neuronal death

JNKs are implicated in a range of brain pathologies and receive considerable attention as potential therapeutic targets. However, JNKs also regulate physiological and homeostatic processes. An attractive hypothesis from the drug development perspective is that distinct JNK isoforms mediate "physiological" and "pathological" responses. However, this lacks experimental evaluation. Here we investigate the isoforms, subcellular pools, and c-Jun/ATF2 targets of JNK in death of central nervous system neurons following withdrawal of trophic support. We use gene knockouts, gene silencing, subcellularly targeted dominant negative constructs, and pharmacological inhibitors. Combined small interfering RNA knockdown of all JNKs 1, 2, and 3, provides substantial neuroprotection. In contrast, knockdown or knock-out of individual JNKs or two JNKs together does not protect. This explains why the evidence for JNK in neuronal death has to date been largely pharmacological. Complete knockdown of c-Jun and ATF2 using small interfering RNA also fails to protect, casting doubt on c-Jun as a critical effector of JNK in neuronal death. Nonetheless, the death requires nuclear but not cytosolic JNK activity as nuclear dominant negative inhibitors of JNK protect, whereas cytosolic inhibitors only block physiological JNK function. Thus any one of the three JNKs is capable of mediating apoptosis and inhibition of nuclear JNK is protective.     

VCP disease associated with myopathy, Paget disease of bone and frontotemporal dementia: review of a unique disorder

Inclusion body myopathy (IBM) associated with Paget disease of the bone (PDB) and frontotemporal dementia (FTD) (now called IBMPFD), is a progressive autosomal dominant disorder that was recently identified as being caused by mutations in the VCP (p97 or CDC48) gene which plays a key role in the ubiquitin-proteasome dependent degradation of cytosolic proteins and in the retro translocation of misfolded proteins from the endoplasmic reticulum into the cytoplasm. Approximately 90% of the affected persons in the study have myopathy or muscle weakness particularly of the shoulder and hip girdles, which can lead to loss of walking ability and even death by complications of respiratory and cardiac failure. About half of affected study participants have Paget disease of bone characterized by abnormal rates of bone growth that can result in bone pain, enlargement and fractures. Findings of premature FTD affecting behavior and personality are seen in a third of affected individuals. Within 20 IBMPFD families whose data was analyzed for this study, ten missense mutations have been identified, the majority of which are located in the N-terminal ubiquitin binding domain. Inclusions seen in the muscle, brain and heart in VCP disease contain ubiquitin, beta amyloid and TDP-43, also seen in other neurodegenerative disorders thus implicating common pathways in their pathogenesis.     

Ability to assess nest predation risk in secondary hole-nesting birds: an experimental study

Because nest predation is the major source of nesting mortality in birds, site-specific predation risk may play an important role in determining birds' ability to select nest sites that reduce predation risk. This possibility has not been adequately tested. Here we report on 5-year experiments by which we studied, independently from birds' earlier experience with specific nest boxes, both the selection and predation risk of nest sites in the common goldeneye (Bucephala clangula). New, previously unoccupied nest boxes were erected in two habitat types on three study areas. Experimentally measured predation risk in the nest boxes varied between 0 and 1.0, i.e. goldeneye females could select a nest site along a wide gradient of possible predation-risk values. We did not find a difference in predation risk between occupied and unoccupied nest boxes, nor was the order of nest box occupation associated with predation risk. A power analysis revealed that our test had reasonably high power to reject a false null hypothesis. Our results suggest that common goldeneye females likely have not evolved an ability to assess predation risk of new, previously unoccupied nest sites.     

Costs and benefits of female-biased natal philopatry in the common goldeneye

Sex-biased natal dispersal in long-lived species may resultin interactions between parents and mature young of the philopatricsex. To investigate the evolutionary basis of natal philopatryin a noncooperative species, the common goldeneye Bucephalaclangula, we studied possible costs and benefits of simultaneousbreeding of females and philopatric daughters. We did not find any fitness consequences of a daughter's breeding on their mother'sbreeding in terms of nest-site selection, body weight, clutchsize, hatching date, or hatching success. Our results, therefore,did not support the assumption of the local resource competitionhypothesis, that the natally philopatric sex should be morecostly to a breeding parent. As possible benefits for daughters returning to their natal area, we tested inheritance of nestsites from mothers and explored whether daughters utilize thepresence of their mother by parasitically sneaking into hermother's nest. Daughters' nest-site selection was not associatedwith the presence of their mothers. A comparison between daughtersand control females revealed that daughters chose their nestsite closer to their natal nest than expected by nest-siteavailability alone. Daughters could not expect to inherit anest site from their mother, and we did not find other indicationsof cooperation between relatives either. The mother's clutchsize did not increase in the year breeding with the daughter, indicating daughters do not parasitize their mother's nest.We suggest that benefits such as decreased nest predation riskassociated with nesting close to the natal nest site may beimportant in the natal philopatric behavior of the species.     

Vertical and seasonal distributions of micro-organisms,zooplankton and phytoplankton in a eutrophic lake

The vertical distributions of bacteria, picoalgae,protozoan and metazoan zooplankton, and phytoplanktonin the highly eutrophic Lake Köyliönjärvi(SW Finland) were studied monthly during the period ofice-cover in January-April 1996. For comparison, wealso provide some data on the distributions of theplankton during the summer. The whole watercolumn remained oxic during the ice-covered period,although the near-bottom oxygen concentrations werealways very low. The heterotrophic nanoflagellateswere more abundant in winter than in summer, butciliates, picoalgae and bacteria were more numerous insummer. In general both zooplankton and phytoplanktonhad low biomass during the ice-covered period.However, the biomass of the diatom Aulacoseiraislandica ssp. islandica was high under the icein April. The calanoid copepod Eudiaptomusgraciloides was the dominant zooplankton species fromJanuary to March, but had almost disappeared by thebeginning of April and did not increase again until inJune. The dominant rotifer species in winterwere Keratella cochlearis, Filinia terminalis,and Filinia longiseta in the surface water andRotaria neptunia near the bottom.     

Preparative variant of the purification of a trypsin-like thrombolytic enzyme from Actinomyces 771

A simple large-scale purification scheme for a trypsin-like enzyme from Actinomyces 771 was developed using carboxylic cation exchange resin Soloze K and DEAE-cellulose. The electrophoretically homogeneous enzyme with the specific trypsin activity of 1.4 U/mg was obtained with a yield of 53%.     

In vitro wear simulation on the RandomPOD wear testing system as a screening method for bearing materials intended for total knee arthroplasty

The 16-station RandomPOD wear test system, previously validated for prosthetic hip wear, was used in the simulation of knee wear mechanisms with a ball-on-flat test configuration. This consisted of a CoCr pin with a ground and polished spherical bearing surface (radius 28 mm) against a conventional, gamma-sterilized UHMWPE disk in serum lubrication. The biaxial motion, consisting of x and y translations, and the load was non-cyclic. Relative to the disk, the center of contact wandered within a circle of 10 mm diameter, and the average sliding velocity was 15.5 mm/s (ranging from 0 to 31 mm/s). The load varied non-cyclically between 0 and 142 N (average 73 N). In the 60-day test with 16 similar wear couples, moderate adhesive wear, the principal wear mechanism of a well-functioning prosthetic knee, dominated. This showed as a burnished, circular wear mark (diameter 13.2 mm, area 137 mm²). The wear factor was 2.04±0.03×10⁻⁶ mm³/N m (mean±95 percent confidence limit). For the first time a truly multidirectional, realistic and uniform, large capacity pin-on-disk simulation of knee wear mechanisms was implemented.     

Correction to: Genetic characterization of Addison’s disease in Bearded Collies

Diversity of the CRISPR locus of Mycobacterium tuberculosis complex has been studied since 1997 for molecular epidemiology purposes. By targeting solely the 43 spacers present in the two first sequenced genomes (H37Rv and BCG), it gave a biased idea of CRISPR diversity and ignored diversity in the neighbouring cas-genes. We set up tailored pipelines to explore the diversity of CRISPR-cas locus in Short Reads. We analyzed data from a representative set of 198 clinical isolates as evidenced by well-characterized SNPs. We found a relatively low diversity in terms of spacers: we recovered only the 68 spacers that had been described in 2000. We found no partial or global inversions in the sequences, letting always the Direct Variant Repeats (DVR) in the same order. In contrast, we found an unexpected diversity in the form of: SNPs in spacers and in Direct Repeats, duplications of various length, and insertions at various locations of the IS6110 insertion sequence, as well as blocks of DVR deletions. The diversity was in part specific to lineages. When reconstructing evolutionary steps of the locus, we found no evidence for SNP reversal. DVR deletions were linked to recombination between IS6110 insertions or between Direct Repeats. This work definitively shows that CRISPR locus of M. tuberculosis did not evolve by classical CRISPR adaptation (incorporation of new spacers) since the last most recent common ancestor of virulent lineages. The evolutionary mechanisms that we discovered could be involved in bacterial adaptation but in a way that remains to be identified.