Early weaning increases aggression and stereotypic behaviour in cats

Behaviour problems are common in companion felines, and problematic behaviour may be a sign of chronic stress. In laboratory animals, early weaning increases the risk for aggression, anxiety, and stereotypic behaviour. However, very few studies have focused on early weaning in one of the world's most popular pets, the domestic cat, although weaning soon after the critical period of socialisation is common practice. To study the effects of early weaning (<12 weeks) on behaviour, a large data set (N = 5726, 40 breeds) was collected from home-living domestic cats through a questionnaire survey. The results show that weaning before 8 weeks of age increases the risk for aggression, but not fearful behaviour. Moreover, cats weaned after 14 weeks of age have a lower probability for aggression towards strangers than early weaned cats and a lower probability for stereotypic behaviour (excessive grooming) than cats weaned at 12 weeks. The effect of weaning age on stereotypic behaviour is partially explained by the effects on aggression. These findings indicate that early weaning has a detrimental effect on behaviour, and suggest delayed weaning as a simple and inexpensive approach to significantly improve the welfare of millions of domestic cats.Peer reviewe

Purification of Highly Active Alphavirus Replication Complexes Demonstrates Altered Fractionation of Multiple Cellular Membranes

Positive-strand RNA viruses replicate their genomes in membrane-associated structures; alphaviruses and many other groups induce membrane invaginations called spherules. Here, we established a protocol to purify these membranous replication complexes (RCs) from cells infected with Semliki Forest virus (SFV). We isolated SFV spherules located on the plasma membrane and further purified them using two consecutive density gradients. This revealed that SFV infection strongly modifies cellular membranes. We removed soluble proteins, the Golgi membranes, and most of the mitochondria, but plasma membrane, endoplasmic reticulum (ER), and late endosome markers were retained in the membrane fraction that contained viral RNA synthesizing activity, replicase proteins, and minus-and plus-strand RNA. Electron microscopy revealed that the purified membranes displayed spherule-like structures with a narrow neck. This membrane enrichment was specific to viral replication, as such a distribution of membrane markers was only observed after infection. Besides the plasma membrane, SFV infection remodeled the ER, and the cofractionation of the RC-carrying plasma membrane and ER suggests that SFV recruits ER proteins or membrane to the site of replication. The purified RCs were highly active in synthesizing both genomic and subgenomic RNA. Detergent solubilization destroyed the replication activity, demonstrating that the membrane association of the complex is essential. Most of the newly made RNA was in double-stranded replicative molecules, but the purified complexes also produced single-stranded RNA as well as released newly made RNA. This indicates that the purification established here maintained the functionality of RCs and thus enables further structural and functional studies of active RCs. IMPORTANCE Similar to all positive-strand RNA viruses, the arthropod-borne alpha-viruses induce membranous genome factories, but little is known about the arrangement of viral replicase proteins and the presence of host proteins in these replication complexes. To improve our knowledge of alphavirus RNA-synthesizing complexes, we isolated and purified them from infected mammalian cells. Detection of viral RNA and in vitro replication assays revealed that these complexes are abundant and highly active when located on the plasma membrane. After multiple purification steps, they remain functional in synthesizing and releasing viral RNA. Besides the plasma membrane, markers for the endoplasmic reticulum and late endosomes were enriched with the replication complexes, demonstrating that alphavirus infection modified cellular membranes beyond inducing replication spherules on the plasma membrane. We have developed here a gentle purification method to obtain large quantities of highly active replication complexes, and similar methods can be applied to other positive-strand RNA viruses.Peer reviewe

The RNA Capping Enzyme Domain in Protein A is Essential for Flock House Virus Replication

The nodavirus flock house virus (FHV) and the alphavirus Semliki Forest virus (SFV) show evolutionarily intriguing similarities in their replication complexes and RNA capping enzymes. In this study, we first established an efficient FHV trans-replication system in mammalian cells, which disjoins protein expression from viral RNA synthesis. Following transfection, FHV replicase protein A was associated with mitochondria, whose outer surface displayed pouch-like invaginations with a ‘neck’ structure opening towards the cytoplasm. In mitochondrial pellets from transfected cells, high-level synthesis of both genomic and subgenomic RNA was detected in vitro and the newly synthesized RNA was of positive polarity. Secondly, we initiated the study of the putative RNA capping enzyme domain in protein A by mutating the conserved amino acids H93, R100, D141, and W215. RNA replication was abolished for all mutants inside cells and in vitro except for W215A, which showed reduced replication. Transfection of capped RNA template did not rescue the replication activity of the mutants. Comparing the efficiency of SFV and FHV trans-replication systems, the FHV system appeared to produce more RNA. Using fluorescent marker proteins, we demonstrated that both systems could replicate in the same cell. This work may facilitate the comparative analysis of FHV and SFV replication.The nodavirus flock house virus (FHV) and the alphavirus Semliki Forest virus (SFV) show evolutionarily intriguing similarities in their replication complexes and RNA capping enzymes. In this study, we first established an efficient FHV trans-replication system in mammalian cells, which disjoins protein expression from viral RNA synthesis. Following transfection, FHV replicase protein A was associated with mitochondria, whose outer surface displayed pouch-like invaginations with a 'neck' structure opening towards the cytoplasm. In mitochondrial pellets from transfected cells, high-level synthesis of both genomic and subgenomic RNA was detected in vitro and the newly synthesized RNA was of positive polarity. Secondly, we initiated the study of the putative RNA capping enzyme domain in protein A by mutating the conserved amino acids H93, R100, D141, and W215. RNA replication was abolished for all mutants inside cells and in vitro except for W215A, which showed reduced replication. Transfection of capped RNA template did not rescue the replication activity of the mutants. Comparing the efficiency of SFV and FHV trans-replication systems, the FHV system appeared to produce more RNA. Using fluorescent marker proteins, we demonstrated that both systems could replicate in the same cell. This work may facilitate the comparative analysis of FHV and SFV replication.Peer reviewe

Effect of Dairy Beef Quality Assurance Training on Dairy Worker Knowledge and Welfare-Related Practices

A study was conducted to determine whether on-farm dairy beef quality assurance (BQA) training affected dairy worker knowledge of BQA and welfare-related practices. Dairy personnel who participated in the BQA training were administered an exam before and after the training to gauge the amount of knowledge gained. The average exam score was 21.0 points higher after the training, increasing from 54.4 to 75.4. Improvement in dairy worker knowledge suggests that BQA training programs have the potential to positively influence the dairy industry through the education of dairy owners and workers on BQA and welfare-related practices

The impact of reference pricing and extension of generic substitution on the daily cost of antipsychotic medication in Finland

Objective: To assess the impact of reference pricing and extension of generic substitution on the daily cost of antipsychotic drugs in Finland during the first year after its launch. Furthermore, the additional impact of reference pricing on prior implemented generic substitution is assessed. Methods: A retrospective analysis was performed between 2006 and 2010. A segmented linear regression analysis of interrupted time series was used to estimate changes in the levels and trends in the cost of one day of treatment. Of the study drugs, clozapine belonged to generic substitution already at the start of the study period while olanzapine and quetiapine were included in generic substitution alongside with reference pricing in 2009. Risperidone was included in generic substitution in 2008, before reference pricing. Results: A substantial decrease in the daily cost of all four antipsychotic substances was seen after one year of the implementation of reference pricing and the extension of generic substitution. The impact ranged from -29.9% to -66.3%, and it was most substantial on the daily cost of olanzapine. Also in the daily cost of risperidone a substantial decrease of -43.3% was observed. However, most of these savings, -32.6%, were generated by generic substitution which had been adopted prior. Conclusions: Reference pricing and the extension of generic substitution produced substantial savings on antipsychotic medication costs during the first year after its launch, but the intensity of the impact differed between active substances. Furthermore, our results suggest that the additional cost savings from reference pricing after prior implemented generic substitution, are comparatively low

Properties of non-structural protein 1 of Semliki Forest virus and its interference with virus replication

Semliki Forest virus (SFV) non-structural protein 1 (nsP1) is a major component of the virus replicase complex. It has previously been studied in cells infected with virus or using transient or stable expression systems. To extend these studies, tetracycline-inducible stable cell lines expressing SFV nsP1 or its palmitoylation-negative mutant (nsP16D) were constructed. The levels of protein expression and the subcellular localization of nsP1 in induced cells were similar to those in virus-infected cells. The nsP1 expressed by stable, inducible cell lines or by SFV-infected HEK293 T-REx cells was a stable protein with a half-life of approximately 5 h. In contrast to SFV infection, induction of nsP1 expression had no detectable effect on cellular transcription, translation or viability. Induction of expression of nsP1 or nsP16D interfered with multiplication of SFV, typically resulting in a 5–10-fold reduction in virus yields. This reduction was not due to a decrease in the number of infected cells, indicating that nsP1 expression does not block virus entry or initiation of replication. Expression of nsP1 interfered with virus genomic RNA synthesis and delayed accumulation of viral subgenomic RNA translation products. Expression of nsP1 with a mutation in the palmitoylation site reduced synthesis of genomic and subgenomic RNAs and their products of translation, and this effect did not resolve with time. These results are in agreement with data published previously, suggesting a role for nsP1 in genomic RNA synthesis

Depression-related work disability: socioeconomic inequalities in onset, duration and recurrence

Objective: Depression is a major cause of disability in working populations and the reduction of socioeconomic inequalities in disability is an important public health challenge. We examined work disability due to depression with four indicators of socioeconomic status. Methods: A prospective cohort study of 125 355 Finnish public sector employees was linked to national register data on work disability (>9 days) due to depressive disorders (International Classification of Diseases, codes F32–F34) from January 2005 to December 2011. Primary outcomes were the onset of work disability due to depressive disorders and, among those with such disability, return to work after and recurrent episodes of work disability due to depression. Results: We found a consistent inverse socioeconomic gradient in work disability due to depression. Lower occupational position, lower educational level, smaller residence size, and rented (vs. owner-occupied) residence were all associated with an increased risk of work disability. Return to work was slower for employees with basic education (cumulative odds ratio = 1.21, 95% CI: 1.05–1.39) compared to those with higher education. Recurrent work disability episodes due to depression were less common among upper-grade non-manual workers (the highest occupational group) than among lower-grade non-manual (hazard ratio = 1.16, 95% CI: 1.07–1.25) and manual (hazard ratio = 1.14, 95% CI: 1.02–1.26) workers. Conclusions: These data from Finnish public sector employees show persistent socioeconomic inequalities in work disability due to depression from 2005 to 2011 in terms of onset, recovery and recurrence

Predictors of employment in young adults with psychiatric work disability

AIM: Mental disorders are the leading cause of work disability among young adults in the industrialized world. Factors predicting employment after long-term psychiatric work disability are largely unknown. METHODS: We linked personal and clinical information from the benefit applications and medical certificates of 1163 young adults (18-34 years) with a new-onset fixed-term psychiatric disability pension in 2008 with employment records between 2005 and 2013. The outcomes were starting employment during and being employed at the end of follow-up. RESULTS: Of the participants, 48% had been employed during and 22% were employed at the end of follow-up. Sustained employment history, university education (master's degree) and no recorded psychological symptoms in childhood were associated with both subsequent employment outcomes. Women and participants under 25 years were more likely to start employment. Depression and other mental disorders (vs psychotic diagnose) and having no comorbid mental disorders or substance abuse were associated with employment at the end of follow-up. CONCLUSIONS: Sustained employment history, university education and no recorded psychological symptoms during childhood predict a return to employment among young adults after a fixed-term psychiatric work disability pension. Pro-active interventions in psychological problems during childhood could enhance employment after a period of work disability

Habitat selection of the mud crab Rhithropanopeus harrisii in its newly invaded range

Information on the habitat selection by non-indigenous species is crucial for understanding their effects on the communities to which they are introduced, since the effects are often focused on the invaded habitats. The North American mud crab Rhithropanopeus harrisii is a new invader in the northern Baltic Sea, on the coasts of Finland and Estonia. In the Finnish Archipelago Sea, it has been found in two very distinct habitats: reed belts of Phragmites australis and algal zones with Fucus vesiculosus as the main habitat-forming species. In previous studies in the Baltic Sea, R. harrisii has preferred F. vesiculosus and has locally driven a shift in the structure of F. vesiculosus-associated invertebrate communities. Here, we disentangled whether habitat choice was determined by habitat structure or the availability of food. First, we conducted a habitat selection experiment with P. australis and F. vesiculosus habitats and varying food availability, and found that R. harrisii preferred F. vesiculosus, with food having no effect on the habitat choice. Second, we studied if the preference for F. vesiculosus was due to the alga itself or the rocks it grows on. We found that R. harrisii preferred the shelter of the rock habitat, indicating that R. harrisii choose their habitat based on habitat structure rather than food availability in the habitat. However, the preference for sheltered rocky bottom habitats also exposes the associated F. vesiculosus communities to the impacts of R. harrisii through predation.Peer reviewe